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Branched Hybridization Chain Reaction Circuit for Ultrasensitive Localizable Imaging of mRNA in Living Cells
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Institute of Chemical Biology and Nanomedicine, State Key Laboratory of Chemo/Bio-Sensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha 410082, P. R. China
*E-mail: [email protected]
*E-mail: [email protected]. Fax: +86-731-88821916.
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Analytical Chemistry

Cite this: Anal. Chem. 2018, 90, 3, 1502–1505
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https://doi.org/10.1021/acs.analchem.7b04848
Published January 4, 2018

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Abstract

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Hybridization chain reaction (HCR) circuits are valuable approaches to monitor low-abundance mRNA, and current HCR is still subjected to issues such as limited amplification efficiency, compromised localization resolution, or complicated designs. We report a novel branched HCR (bHCR) circuit for efficient signal-amplified imaging of mRNA in living cells. The bHCR can be realized using a simplified design by hierarchically coupling two HCR circuits with two split initiator fragments of the secondary HCR circuit incorporated in the probes for the primary HCR circuit. The bHCR circuit enables one to generate a hyperbranched assembly seeded from a single target initiator, affording the potential for localizing single target molecules in live cells. In vitro assays show that bHCR offers very high amplification efficiency and specificity in single mismatch discrimination with a detection limit of 500 fM. Live cell studies reveal that bHCR displays intense fluorescence spots indicating mRNA localization in living cells with improved contrast. The bHCR method can provide a useful platform for low-abundance biomarker detection and imaging for cell biology and diagnostics.

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Messenger RNAs (mRNAs) play a fundamental role in conveying the genetic blueprint encoded by DNA to proteins. Abnormalities of mRNA expression could be useful biomarkers for various diseases such as cancer. (1) Hybridization based probes allowing directly detection and imaging of endogenous mRNA in living cells afford a valuable approach for study of mRNA biology and diagnosis of related diseases. (2) Molecular beacons (MBs) (3, 4) and spherical nucleic acids (SNAs) (5, 6) are the most common probes for mRNA detection in living cells. These probes convert the mRNA hybridization events into fluorescence signals in an equivalent reaction ratio. The lack of signal amplification in these probes limits their utility for expression analysis of low-abundance mRNA. While expression of mRNA is heterogeneous, failures in monitoring the low-abundance subpopulations could be impediments to understanding the functions of these mRNA and early assessment of the associated diseases. (7)

Nucleic acid amplification provides a direct solution to expression analysis of low-abundance mRNA. (8, 9) Recent development of nucleic acid circuits, such as hybridization chain reaction (HCR), (10-12) cascade hybridization reaction, (13) catalyzed hairpin assembly (CHA), (14) and entropy driven catalysis, (15) has enabled programming of the kinetically controlled assembly of DNA modules, which creates an efficient approach for nonenzymatic amplification. An obvious advantage of such isothermal, enzyme-free amplification is the applicability to highly sensitive nucleic acid based sensors for signal amplified imaging in living cells. Motivated by this rationale, others (14, 16, 17) and our group (18) have developed sensitive imaging approaches for visualizing RNAs in living cells. Nonetheless, current nonenzymatic amplification circuits are realized in chain-like reactions, only conferring limited efficiency in signal amplification and compromised resolution for localization. To address this issue, new HCR circuits with nonlinear or branched reactions have been developed. (19-22) We have demonstrated a branched HCR design that is realized using multistep consecutive reactions for visualization of single mRNA molecule in fixed cells or tissues. (19) However, the multiple reaction and washing steps preclude its applications in living cells. Other dendritic or hyperbranched HCR circuits, though allowing generating a hyperbranched assembly in a single reaction, (20-23) still require complicated designs using multiple-hairpin probes or multiple probes. Moreover, these nonlinear HCR circuits have been largely unexplored for mRNA imaging.

Herein, we report a novel branched HCR (bHCR) circuit that allows formation of a hyperbranched assembly of probes in a single reaction using a simplified design for efficient signal-amplified imaging of mRNA in living cells. This bHCR strategy relies on a hierarchical coupling design of two HCR circuits in a single reaction, as illustrated in Scheme 1. A primary linear HCR circuit is designed using two hairpin probes H1 and H2, which are triggered by target mRNA to form the backbone chain of the hyperbranched assembly. A distinct design of probes H1 and H2 is that two split initiator fragments of the secondary HCR are incorporated in the loop region of H1 and the toehold region of H2, respectively. When the backbone assembly forms in response to target mRNA, these split initiator fragments are drawn into close proximity, activating the secondary HCR circuit and enabling branched growth of chain-like assembly between hairpin probes H3 and H4. The hierarchical coupling of the primary and the secondary HCR circuits is able to generate a hyperbranched, brush-like assembly with a single mRNA target as the initiator. In order to light up the mRNA target, the probe F–H3 is designed to have a fluorophore FAM and a quencher BHQ1 in the stem region of H3. This design allows the probe to exhibit a low fluorescence background in its folded, hairpin state while deliver enhanced fluorescence in the hyperbranched assembly in which H3 is extended by hybridization with H4. When the probes H1, H2, H3, and H4 are transfected into cells, a hyperbranched, highly fluorescent assembly of these probes is generated seeded from each single mRNA target, enabling high-contrast and spatially localizable imaging of the mRNA target. Hence, our design can provide a useful platform for highly sensitive and localizable imaging of intracellular mRNA, especially for the low-abundance subpopulations.

Scheme 1

Scheme 1. Working Principle of bHCR for mRNA Imaging
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To demonstrate the principle of our bHCR strategy for mRNA imaging in living cells, we chose survivin mRNA, an important biomarker overexpressed in many malignant tumors, (24) as the model target. On the basis of bioinformatic calculation of the secondary structure of the mRNA, a target region of 24 nucleotides with no predicted hairpin structure was selected. Accordingly, the hairpin probes for the primary HCR circuit H1 and H2 and for the secondary circuit H3 and H4 were designed. To investigate the feasibility of these probes, the primary and secondary HCR circuits were first analyzed using agarose gel electrophoresis. Gel images showed that bright ladder-like bands characteristic for HCR circuits were obtained when 1 μM target RNA sequence T was incubating with 1 μM H1 and 1 μM H2, or 1 μM secondary initiator sequence T2 was incubating with 1 μM H3 and 1 μM H4 (Figure S-1). The results verified HCR assembly of H1 and H2 or H3 and H4 seeded by target RNA sequence or secondary initiator sequence, respectively. We also observed brighter bands with larger molecular weights with lower concentration of the initiators, suggesting a high ratio of probes to initiator gave larger HCR assembly. No new bands appeared when the initiator was absent. These results evidenced successful construction of the primary and secondary HCR circuits. After confirming two individual HCR circuits, we examined bHCR using gel electrophoresis (Figure 1a). After incubating 250 nM target T with 500 nM H1, 500 nM H2, 1 μM H3, and 1 μM H4, we obtained many bright bands with size around 10 000 bp (lane 4). In contrast, the linear HCR products obtained by incubating 250 nM T with 500 nM H1, 500 nM H2 (lane 1), or incubating 500 nM T2 with 1 μM H3 and 1 μM H4 (lane 2) gave much smaller sizes. Incubation of 250 nM T with 500 nM H1 and 500 nM H2 (lane 3) and 1 μM H3 also gave smaller-size bands as linear HCR. These observations manifested that coupling of two individual HCR circuits afforded larger assembly of the probes, validating the proposed bHCR design. In the absence of target T, there were some low-fluorescence bands (lane 5), which were attributed to marginal system leakage from imperfectly annealed hairpins.

Figure 1

Figure 1. (a) Gel electrophoresis analysis of bHCR: lane 1, 500 nM H1 and 500 nM H2 with 250 nM T; lane 2, 1 μM H3 and 1 μM H4 with 500 nM T2; lane 3, 500 nM H1, 500 nM H2, 1 μM H3 with 250 nM T; lane 4, 500 nM H1, 500 nM H2, 1 μM H3 and 1 μM H4 with 250 nM T; lane 5, 500 nM H1, 500 nM H2, 1 μM H3 and 1 μM H4. (b–f) AFM images of the assembly products in primary HCR circuit (b), secondary HCR circuit (c), bHCR system in the presence of T (d), and absence of T (e). Part f is enlarged details of part d. Scale bar: 200 nm.

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To further confirm the bHCR system, we directly visualized the morphology of the assembly products with AFM imaging. When only incubating H1, H2, H3, and H4, we obtained tiny spots with heights of ∼1.5 nm (Figure 1e), which were ascribed to the hairpin probes failing to be assembled in the absence of target RNA. In contrast, linear structures were found when T was incubated with H1 and H2 or T2 was incubated with H3 and H4 (Figure 1b,c), suggesting linear polymers generated in HCR. Many branched structures were found in the bHCR system upon incubating target T with H1, H2, H3, and H4 (Figure 1e,f and Figure S-2). The bHCR products were polydisperse both in sizes and branching efficiency because of the random assembly of the hairpins which, therefore, also generated a few smaller linear polymers. These images gave clear evidence for branched assembly in the bHCR system. This finding was consistent with the extended bands of varying molecular weights in the gel images.

The bHCR system was further investigated using fluorescence spectroscopy for in vitro detection of target sequence. Incubation of 200 nM H1, 200 nM H2, 200 nM F–H3, and 200 nM H4 for 3 h at 37 °C only gave a weak fluorescence peak (Figure 2a), indicating low fluorescence background and marginal leakage for the bHCR system. Appreciable fluorescence activation was observed when 2 nM target sequence T was incubated with 200 nM H1, 200 nM H2, 200 nM F–H3, and 200 nM H4, and the signal-to-background ratio was ∼8 fold. This observation implied high-contrast activation of the bHCR system in response to the target. Interestingly, incubation of 200 nM F–H1 and 200 nM H2 with 2 nM T merely showed a slight increase of the fluorescence peak, and the signal-to-background ratio (4.4-fold) was much smaller than that obtained in the bHCR system. This result clearly demonstrated the higher amplification efficiency of bHCR than conventional linear HCR. Selectivity analysis of the bHCR systems was examined using a one-base mutation sequence of the target. The obtained fluorescence intensity exhibited little enhancement to the background, implying that the bHCR system afforded very high selectivity with the ability to discriminating single-base mutations. The fluorescence peaks were also found to be dynamically dependent upon the concentrations of target sequence in a three-decade range from 1 pM to 2 nM (Figure 2b). A quasi-linear correlation was obtained for peak intensities at 523 nm to target concentrations ranging from 1 pM to 0.8 nM (Figure 2c). The detection limit was estimated as low as 500 fM, which was much better than current dendritic or hyperbranched HCR circuits. (20-23) A further comparison was performed between the bHCR system and conventional linear HCR circuit consisting of hairpins F–H1 and H2 and the corresponding initiator T. It was found that the fluorescence background of the linear HCR system was 2-fold lower than that of the bHCR circuit. The result was sensible as the system leakage was also amplified by the hierarchical coupling of two HCR circuits. The fluorescence responses of the linear HCR system were also dynamically correlated to target concentrations, and the peak fluorescence intensities showed linear correlation to target concentrations in the range from 20 pM to 5 nM with a detection limit of 10 pM (Figure S-1). The results revealed that the bHCR system offered higher sensitivity than the linear HCR assay. At different target concentrations, the bHCR system was found to consistently give higher signal-to-background ratios than the linear HCR (Figure 2d). These data testified that bHCR provided much higher amplification efficiency than the linear HCR system, indicating the successful branched growth of the brush-like secondary HCR polymers along the primary HCR assemblies. In addition, time-dependent measurements showed that the peak intensities increased with increasing reaction time, and at a lower concentration the reaction required prolonged time to achieved saturated responses (Figure S-4).

Figure 2

Figure 2. (a) Fluorescence spectral responses for mixtures of 200 nM H1 and 200 nM H2 (blue); 200 nM F–H1 and 200 nM H2 with 2 nM T (green); 200 nM H1 and 200 nM H2, 200 nM F–H3 and 200 nM H4 (black); 200 nM H1, 200 nM H2, 200 nM F–H3, and 200 nM H4 with 2 nM T (pink); or with 2 nM one-base mismatched T (Mu-T) (red). (b) Fluorescence responses of bHCR system to T of varying concentrations. (c) Calibration curves of fluorescence intensities versus target concentrations. Inset shows the linear relationship between fluorescence intensities and target concentrations in the linear range. (d) Histograms of signal-to-background (STB) ratios of bHCR system (blue) and linear system (red) at different target concentrations.

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Next, we explored the ability of bHCR for imaging of mRNA in living cells. When HeLa cells were transfected with the hairpin probes H1, H2, F–H3, and H4 using Lipofectamine 3000, very intense green fluorescent spots was observed in the cells (Figure 3a). A control experiment using C166 cells with no expression of target mRNA (25) revealed that almost no fluorescent spots appeared in the cells (Figure 3b). This result evidenced that these bright fluorescent spots found in HeLa cells were specific to target mRNA, implying that fluorescent spots were ascribed to the hyperbranched assembly of hairpin probes seeded from target mRNA. Furthermore, we transfected HeLa cells using only two probes F–H1 and H2 in the linear HCR system. As anticipated, the image only showed fewer and much weak fluorescent spots in the cells. The average fluorescence intensity of the spots in the bHCR circuit were found to be ∼5-fold enhancement as compared to that for the linear HCR system. Further flow cytometry assay of these three types of cells also revealed that bHCR gave much higher fluorescence signals than linear HCR (Figure S-5). These findings clearly proved that bHCR had the promise in affording higher amplification efficiency and better sensitivity in live cell imaging relative to linear HCR.

Figure 3

Figure 3. Fluorescence images for (a) HeLa cells transfected with H1, H2, F–H3, and H4; (b) C166 cells transfected with H1, H2, and F–H3 H4; (c) HeLa cells transfected with F–H1 and H2. (1) Fluorescence; (2) merged with DIC. Scale bar: 20 μm.

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The bHCR circuits was further applied to fluorescence imaging of different cell lines, HeLa, HepG-2, and C166. It was found that HeLa cells showed many more fluorescence spots than HepG2 cells (Figure S-6), indicating higher expression of survivin mRNA in HeLa cells than HepG2 cells. This result was consistent with the finding reported previously. (25) With HeLa cells pretreated using 5 nM or 10 nM YM155, an imidazolium-based compound specifically repressing survivin mRNA expression, (26) followed by transfection of the probes, we also found quite a few intense fluorescent spots in the cells treated using 5 nM YM155 and much less fluorescent spots appeared in the cells treated using 5 nM YM155 (Figure S-7). These results confirmed the knock-down effect of YM155 in regulating survivin mRNA. To validate the results, quantitative reverse transcription polymerase chain reaction (RT-qPCR) analysis of the expression of survivin mRNA in these cells was performed (Figure S-8). The results revealed that the number of intense fluorescent spots in the cells increased with increasing expression levels of target mRNA in the cells. This result verified the ability of the bHCR circuit for quantitative imaging mRNA expression in living cells.

In conclusion, we successfully demonstrated a novel bHCR circuit for ultrasensitive imaging of mRNA in living cells. This bHCR strategy exploited a hierarchical coupling of two HCR circuits in a single reaction and could be developed using a simplified design by incorporating two split initiator fragments in the probes in the primary HCR circuit. The bHCR circuit enabled to generated a hyperbranched assembly seeded from a single target initiator, which provided the possibility of localizing single target molecules in live cell imaging applications. The result revealed that bHCR generated highly branched structures with high molecular weights in response to target sequence with a detection limit of 500 fM and high specificity in single mismatch discrimination. The bHCR system was also demonstrated to display intense fluorescence spots indicating the localization of target mRNA in living cells. Collectively, this bHCR strategy can provide a powerful platform for low-abundance biomarker detection and imaging for cell biology and clinical diagnostics.

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The Supporting Information is available free of charge on the ACS Publications website at DOI: 10.1021/acs.analchem.7b04848.

  • Experimental methods including materials and instruments, gel electrophoresis analysis, AFM imaging, fluorescence measurements, cell culture and fluorescence imaging, flow cytometry assay, and RT-PCR assay (PDF)

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Author Information

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  • Corresponding Authors
    • Ru-Qin Yu - Institute of Chemical Biology and Nanomedicine, State Key Laboratory of Chemo/Bio-Sensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha 410082, P. R. China;  Orcidhttp://orcid.org/0000-0002-7412-8360;  Email: [email protected]
    • Jian-Hui Jiang - Institute of Chemical Biology and Nanomedicine, State Key Laboratory of Chemo/Bio-Sensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha 410082, P. R. China;  Orcidhttp://orcid.org/0000-0003-1594-4023;  Email: [email protected]
  • Authors
    • Lan Liu - Institute of Chemical Biology and Nanomedicine, State Key Laboratory of Chemo/Bio-Sensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha 410082, P. R. China
    • Jin-Wen Liu - Institute of Chemical Biology and Nanomedicine, State Key Laboratory of Chemo/Bio-Sensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha 410082, P. R. China
    • Han Wu - Institute of Chemical Biology and Nanomedicine, State Key Laboratory of Chemo/Bio-Sensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha 410082, P. R. China
    • Xiang-Nan Wang - Institute of Chemical Biology and Nanomedicine, State Key Laboratory of Chemo/Bio-Sensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha 410082, P. R. China
  • Notes
    The authors declare no competing financial interest.

Acknowledgment

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This work was supported by NSFC (Grants 21527810 and 21521063).

References

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This article references 26 other publications.

  1. 1
    Tyagi, S. Nat. Methods 2009, 6, 331– 338 DOI: 10.1038/nmeth.1321
    Google Scholar
    1
    Imaging intracellular RNA distribution and dynamics in living cells
    Tyagi, Sanjay
    Nature Methods (2009), 6 (5), 331-338CODEN: NMAEA3; ISSN:1548-7091. (Nature Publishing Group)
    A review. Powerful methods now allow the imaging of specific mRNAs in living cells. These methods enlist fluorescent proteins to illuminate mRNAs, use labeled oligonucleotide probes and exploit aptamers that render org. dyes fluorescent. The intracellular dynamics of mRNA synthesis, transport and localization can be analyzed at higher temporal resoln. with these methods than has been possible with traditional fixed-cell or biochem. approaches. These methods have also been adopted to visualize and track single mRNA mols. in real time. This review explores the promises and limitations of these methods.
  2. 2
    Xia, Y.; Zhang, R.; Wang, Z.; Tian, J.; Chen, X. Chem. Soc. Rev. 2017, 46, 2824– 2843 DOI: 10.1039/C6CS00675B
    Google Scholar
    2
    Recent advances in high-performance fluorescent and bioluminescent RNA imaging probes
    Xia, Yuqiong; Zhang, Ruili; Wang, Zhongliang; Tian, Jie; Chen, Xiaoyuan
    Chemical Society Reviews (2017), 46 (10), 2824-2843CODEN: CSRVBR; ISSN:0306-0012. (Royal Society of Chemistry)
    RNA plays an important role in life processes. Imaging of mRNAs (mRNAs) and micro-RNAs (miRNAs) not only allows us to learn the formation and transcription of mRNAs and the biogenesis of miRNAs involved in various life processes, but also helps in detecting cancer. High-performance RNA imaging probes greatly expand our view of life processes and enhance the cancer detection accuracy. In this review, we summarize the state-of-the-art high-performance RNA imaging probes, including exogenous probes that can image RNA sequences with special modification and endogeneous probes that can directly image endogenous RNAs without special treatment. For each probe, we review its structure and imaging principle in detail. Finally, we summarize the application of mRNA and miRNA imaging probes in studying life processes as well as in detecting cancer. By correlating the structures and principles of various probes with their practical uses, we compare different RNA imaging probes and offer guidance for better utilization of the current imaging probes and the future design of higher-performance RNA imaging probes.
  3. 3
    Tyagi, S.; Kramer, F. R. Nat. Biotechnol. 1996, 14, 303– 308 DOI: 10.1038/nbt0396-303
    Google Scholar
    3
    Molecular beacons: probes that fluoresce upon hybridization
    Tyagi, Sanjay; Kramer, Fred Russell
    Nature Biotechnology (1996), 14 (3), 303-8CODEN: NABIF9; ISSN:1087-0156. (Nature Publishing Co.)
    The authors have developed novel nucleic acid probes that recognize and report the presence of specific nucleic acids in homogeneous solns. These probes undergo a spontaneous fluorogenic conformational change when they hybridize to their targets. Only perfectly complementary targets elicit this response, as hybridization does not occur when the target contains a mismatched nucleotide or a deletion. The probes are particularly suited for monitoring the synthesis of specific nucleic acids in real time. When used in nucleic acid amplification assays, gene detection is homogeneous and sensitive, and can be carried out in a sealed tube. When introduced into living cells, these probes should enable the origin, movement, and fate of specific mRNAs to be traced.
  4. 4
    Qiu, L.; Wu, C.; You, M.; Han, D.; Chen, T.; Zhu, G.; Jiang, J.; Yu, R.; Tan, W. J. Am. Chem. Soc. 2013, 135, 12952– 12955 DOI: 10.1021/ja406252w
    Google Scholar
    4
    A Targeted, Self-Delivered, and Photocontrolled Molecular Beacon for mRNA Detection in Living Cells
    Qiu, Liping; Wu, Cuichen; You, Mingxu; Han, Da; Chen, Tao; Zhu, Guizhi; Jiang, Jianhui; Yu, Ruqin; Tan, Weihong
    Journal of the American Chemical Society (2013), 135 (35), 12952-12955CODEN: JACSAT; ISSN:0002-7863. (American Chemical Society)
    The spatiotemporal dynamics of specific mRNA mols. are difficult to image and detect inside living cells, and this has been a significant challenge for the chem. and biomedical communities. To solve this problem, we have developed a targeted, self-delivered, and photocontrolled aptamer-based mol. beacon (MB) for intracellular mRNA anal. An internalizing aptamer connected via a double-stranded DNA structure was used as a carrier probe (CP) for cell-specific delivery of the MB designed to signal target mRNA. A light activation strategy was employed by inserting two photolabile groups in the CP sequence, enabling control over the MB's intracellular function. After the probe was guided to the target cell via specific binding of aptamer AS1411 to nucleolin on the cell membrane, light illumination released the MB for mRNA monitoring. Consequently, the MB is able to perform live-cell mRNA imaging with precise spatiotemporal control, while the CP acts as both a tracer for intracellular distribution of the MB before photoinitiation and an internal ref. for mRNA ratiometric detection.
  5. 5
    Seferos, D. S.; Giljohann, D. A.; Hill, H. D.; Prigodich, A. E.; Mirkin, C. A. J. Am. Chem. Soc. 2007, 129, 15477– 15479 DOI: 10.1021/ja0776529
    Google Scholar
    5
    Nano-Flares: Probes for Transfection and mRNA Detection in Living Cells
    Seferos, Dwight S.; Giljohann, David A.; Hill, Haley D.; Prigodich, Andrew E.; Mirkin, Chad A.
    Journal of the American Chemical Society (2007), 129 (50), 15477-15479CODEN: JACSAT; ISSN:0002-7863. (American Chemical Society)
    We demonstrate that novel oligonucleotide-modified gold nanoparticle probes hybridized to fluorophore-labeled complements can be used as both transfection agents and cellular "nano-flares" for detecting mRNA in living cells. Nano-flares take advantage of the highly efficient fluorescence quenching properties of gold, cellular uptake of oligonucleotide nanoparticle conjugates without the use of transfection agents, and the enzymic stability of such conjugates, thus overcoming many of the challenges to creating sensitive and effective intracellular probes. Nano-flares exhibit high signaling, have low background fluorescence, and are sensitive to changes in the no. of RNA transcripts present in cells.
  6. 6
    Cutler, J. I.; Auyeung, E.; Mirkin, C. A. J. Am. Chem. Soc. 2012, 134, 1376– 1391 DOI: 10.1021/ja209351u
    Google Scholar
    6
    Spherical Nucleic Acids
    Cutler, Joshua I.; Auyeung, Evelyn; Mirkin, Chad A.
    Journal of the American Chemical Society (2012), 134 (3), 1376-1391CODEN: JACSAT; ISSN:0002-7863. (American Chemical Society)
    A review. A historical perspective of the development of spherical nucleic acid (SNA) conjugates and other three-dimensional nucleic acid nanostructures is provided. This Perspective details the synthetic methods for prepg. them, followed by a discussion of their unique properties and theor. and exptl. models for understanding them. Important examples of technol. advances made possible by their fundamental properties spanning the fields of chem., mol. diagnostics, gene regulation, medicine, and materials science are also presented.
  7. 7
    Visvader, J. E. Nature 2011, 469, 314– 322 DOI: 10.1038/nature09781
    Google Scholar
    There is no corresponding record for this reference.
  8. 8
    Zhang, H.; Li, F.; Dever, B.; Li, X.; Le, X. C. Chem. Rev. 2013, 113, 2812– 2841 DOI: 10.1021/cr300340p
    Google Scholar
    8
    DNA-Mediated Homogeneous Binding Assays for Nucleic Acids and Proteins
    Zhang, Hongquan; Li, Feng; Dever, Brittany; Li, Xing-Fang; Le, X. Chris
    Chemical Reviews (Washington, DC, United States) (2013), 113 (4), 2812-2841CODEN: CHREAY; ISSN:0009-2665. (American Chemical Society)
    A review. The present review mainly focuses on recent advances in aptamer beacons, catalytic beacons, in vitro isothermal amplification techniques, DNA nanostructures, and binding-induced DNA assembly for the detection of specific nucleic acid sequences and proteins.
  9. 9
    Zhao, Y.; Chen, F.; Li, Q.; Wang, L.; Fan, C. Chem. Rev. 2015, 115, 12491– 12545 DOI: 10.1021/acs.chemrev.5b00428
    Google Scholar
    9
    Isothermal Amplification of Nucleic Acids
    Zhao, Yongxi; Chen, Feng; Li, Qian; Wang, Lihua; Fan, Chunhai
    Chemical Reviews (Washington, DC, United States) (2015), 115 (22), 12491-12545CODEN: CHREAY; ISSN:0009-2665. (American Chemical Society)
    Isothermal amplification of nucleic acids is a simple process that rapidly and efficiently accumulates nucleic acid sequences at const. temp. Since the early 1990s, various isothermal amplification techniques have been developed as alternatives to polymerase chain reaction (PCR). These isothermal amplification methods have been used for biosensing targets such as DNA, RNA, cells, proteins, small mols., and ions. The applications of these techniques for in situ or intracellular bioimaging and sequencing have been amply demonstrated. Amplicons produced by isothermal amplification methods have also been utilized to construct versatile nucleic acid nanomaterials for promising applications in biomedicine, bioimaging, and biosensing. The integration of isothermal amplification into microsystems or portable devices improves nucleic acid-based on-site assays and confers high sensitivity. Single-cell and single-mol. analyses have also been implemented based on integrated microfluidic systems. In this review, we provide a comprehensive overview of the isothermal amplification of nucleic acids encompassing work published in the past two decades. First, different isothermal amplification techniques are classified into three types based on reaction kinetics. Then, we summarize the applications of isothermal amplification in bioanal., diagnostics, nanotechnol., materials science, and device integration. Finally, several challenges and perspectives in the field are discussed.
  10. 10
    Dirks, R. M.; Pierce, N. A. Proc. Natl. Acad. Sci. U. S. A. 2004, 101, 15275 DOI: 10.1073/pnas.0407024101
    Google Scholar
    10
    Triggered amplification by hybridization chain reaction
    Dirks, Robert M.; Pierce, Niles A.
    Proceedings of the National Academy of Sciences of the United States of America (2004), 101 (43), 15275-15278CODEN: PNASA6; ISSN:0027-8424. (National Academy of Sciences)
    We introduce the concept of hybridization chain reaction (HCR), in which stable DNA monomers assemble only upon exposure to a target DNA fragment. In the simplest version of this process, two stable species of DNA hairpins coexist in soln. until the introduction of initiator strands triggers a cascade of hybridization events that yields nicked double helixes analogous to alternating copolymers. The av. mol. wt. of the HCR products varies inversely with initiator concn. Amplification of more diverse recognition events can be achieved by coupling HCR to aptamer triggers. This functionality allows DNA to act as an amplifying transducer for biosensing applications.
  11. 11
    Bi, S.; Yue, S.; Zhang, S. Chem. Soc. Rev. 2017, 46, 4281– 4298 DOI: 10.1039/C7CS00055C
    Google Scholar
    11
    Hybridization chain reaction: a versatile molecular tool for biosensing, bioimaging, and biomedicine
    Bi, Sai; Yue, Shuzhen; Zhang, Shusheng
    Chemical Society Reviews (2017), 46 (14), 4281-4298CODEN: CSRVBR; ISSN:0306-0012. (Royal Society of Chemistry)
    A review. Developing powerful, simple and low-cost DNA amplification techniques is of great significance to bioanal. and biomedical research. Thus far, many signal amplification strategies have been developed, such as polymerase chain reaction (PCR), rolling circle amplification (RCA), and DNA strand displacement amplification (SDA). In particular, hybridization chain reaction (HCR), a type of toehold-mediated strand displacement (TMSD) reaction, has attracted great interest because of its enzyme-free nature, isothermal conditions, simple protocols, and excellent amplification efficiency. In a typical HCR, an analyte initiates the cross-opening of two DNA hairpins, yielding nicked double helixes that are analogous to alternating copolymers. As an efficient amplification platform, HCR has been utilized for the sensitive detection of a wide variety of analytes, including nucleic acids, proteins, small mols., and cells. In recent years, more complicated sets of monomers have been designed to develop nonlinear HCR, such as branched HCR and even dendritic systems, achieving quadratic and exponential growth mechanisms. In addn., HCR has attracted enormous attention in the fields of bioimaging and biomedicine, including applications in fluorescence in situ hybridization (FISH) imaging, live cell imaging, and targeted drug delivery. In this review, we introduce the fundamentals of HCR and examine the visualization and anal. techniques for HCR products in detail.
  12. 12
    Chen, Y.; Xu, J.; Su, J.; Xiang, Y.; Yuan, R.; Chai, Y. Anal. Chem. 2012, 84, 7750– 7755 DOI: 10.1021/ac3012285
    Google Scholar
    12
    In situ hybridization chain reaction amplification for universal and highly sensitive electrochemiluminescent detection of DNA
    Chen, Ying; Xu, Jin; Su, Jiao; Xiang, Yun; Yuan, Ruo; Chai, Yaqin
    Analytical Chemistry (Washington, DC, United States) (2012), 84 (18), 7750-7755CODEN: ANCHAM; ISSN:0003-2700. (American Chemical Society)
    In this work, we describe a new universal and highly sensitive strategy for electrochemiluminescent (ECL) detection of sequence specific DNA at the femtomolar level via in situ hybridization chain reaction (HCR) signal amplification. The DNA capture probes are self-assembled on a gold electrode. The presence of the target DNA and two hairpin helper DNAs leads to the formation of extended dsDNA polymers through HCR on the electrode surface. The in situ, HCR-generated dsDNA polymers cause the intercalation of numerous ECL indicators (Ru(phen)32+) into the dsDNA grooves, resulting in significantly amplified ECL signal output. The proposed strategy combines the amplification power of the DNA HCR and the inherent high sensitivity of the ECL technique and enables low femtomolar detection of sequence specific DNA. The developed strategy also shows high selectivity against single-base mismatch sequences, which makes our new universal and highly sensitive HCR-based method a useful addn. to the amplified DNA detection arena.
  13. 13
    Cheglakov, Z.; Cronin, T. M.; He, C.; Weizmann, Y. J. Am. Chem. Soc. 2015, 137, 6116– 6119 DOI: 10.1021/jacs.5b01451
    Google Scholar
    13
    Live cell microRNA imaging using cascade hybridization reaction
    Cheglakov, Zoya; Cronin, Timothy M.; He, Chuan; Weizmann, Yossi
    Journal of the American Chemical Society (2015), 137 (19), 6116-6119CODEN: JACSAT; ISSN:0002-7863. (American Chemical Society)
    Recent advances in RNA research have posed new directives in biol. and chem. to uncover the complex roles of ribonucleic acids in cellular processes. Innovative techniques to visualize native RNAs, particularly, short, low-abundance RNAs in live cells, can dramatically impact current research on the roles of RNAs in biol. Herein, we report a novel method for real-time, microRNA imaging inside live cells based on programmable oligonucleotide probes, which self-assemble through the Cascade Hybridization Reaction (CHR).
  14. 14
    Jiang, Y.; Li, B.; Milligan, J. N.; Bhadra, S.; Ellington, A. D. J. Am. Chem. Soc. 2013, 135, 7430 DOI: 10.1021/ja4023978
    Google Scholar
    14
    Real-time detection of isothermal amplification reactions with thermostable catalytic hairpin assembly
    Jiang, Yu; Li, Bingling; Milligan, John N.; Bhadra, Sanchita; Ellington, Andrew D.
    Journal of the American Chemical Society (2013), 135 (20), 7430-7433CODEN: JACSAT; ISSN:0002-7863. (American Chemical Society)
    Catalytic hairpin assembly (CHA) is an enzyme-free amplification method that has previously proven useful in amplifying and transducing signals at the terminus of nucleic acid amplification reactions. Here, for the first time, we engineered CHA to be thermostable from 37 to 60° and in consequence have generalized its application to the real-time detection of isothermal amplification reactions. CHA circuits were designed and optimized for both high- and low-temp. rolling circle amplification (RCA) and strand displacement amplification (SDA). The resulting circuits not only increased the specificity of detection but also improved the sensitivity by as much as 25- to 10000-fold over comparable real-time detection methods. These methods have been condensed into a set of general rules for the design of thermostable CHA circuits with high signals and low noise.
  15. 15
    Zhang, D. Y.; Turberfield, A. J.; Yurke, B.; Winfree, E. Science 2007, 318, 1121– 1125 DOI: 10.1126/science.1148532
    Google Scholar
    15
    Engineering Entropy-Driven Reactions and Networks Catalyzed by DNA
    Zhang, David Yu; Turberfield, Andrew J.; Yurke, Bernard; Winfree, Erik
    Science (Washington, DC, United States) (2007), 318 (5853), 1121-1125CODEN: SCIEAS; ISSN:0036-8075. (American Association for the Advancement of Science)
    Artificial biochem. circuits are likely to play as large a role in biol. engineering as elec. circuits have played in the engineering of electromech. devices. Toward that end, nucleic acids provide a designable substrate for the regulation of biochem. reactions. However, it has been difficult to incorporate signal amplification components. We introduce a design strategy that allows a specified input oligonucleotide to catalyze the release of a specified output oligonucleotide, which in turn can serve as a catalyst for other reactions. This reaction, which is driven forward by the configurational entropy of the released mol., provides an amplifying circuit element that is simple, fast, modular, composable, and robust. We have constructed and characterized several circuits that amplify nucleic acid signals, including a feedforward cascade with quadratic kinetics and a pos. feedback circuit with exponential growth kinetics.
  16. 16
    Wu, C.; Cansiz, S.; Zhang, L.; Teng, I.; Qiu, L.; Li, J.; Liu, Y.; Zhou, C.; Hu, R.; Zhang, T.; Cui, C.; Cui, L.; Tan, W. J. Am. Chem. Soc. 2015, 137, 4900– 4903 DOI: 10.1021/jacs.5b00542
    Google Scholar
    16
    A nonenzymatic hairpin DNA cascade reaction provides high signal gain of mRNA imaging inside live cells
    Wu, Cuichen; Cansiz, Sena; Zhang, Liqin; Teng, I-Ting; Qiu, Liping; Li, Juan; Liu, Yuan; Zhou, Cuisong; Hu, Rong; Zhang, Tao; Cui, Cheng; Cui, Liang; Tan, Weihong
    Journal of the American Chemical Society (2015), 137 (15), 4900-4903CODEN: JACSAT; ISSN:0002-7863. (American Chemical Society)
    Enzyme-free signal amplification has enabled sensitive in vitro detection of biomols. such as proteins and nucleic acids. However, monitoring targets of interest in live cells via enzyme-free amplification is still challenging, esp. for analytes with low concns. To the best of our knowledge, this paper reports the first attempt to perform mRNA imaging inside live cells, using a nonenzymic hairpin DNA cascade reaction for high signal gain, termed a hairpin DNA cascade amplifier (HDCA). In conventional nucleic acid probes, such as linear hybridization probes, mRNA target signaling occurs in an equiv. reaction ratio (1:1), whereas, in HDCA, one mRNA target is able to yield multiple signal outputs (1:m), thus achieving the goal of signal amplification for low-expression mRNA targets. Moreover, the recycled mRNA target in the HDCA serves as a catalyst for the assembly of multiple DNA duplexes, generating the fluorescent signal of reduced MnSOD mRNA expression, thus indicating amplified intracellular imaging. This programmable cascade reaction presents a simple and modular amplification mechanism for intracellular biomarkers of interest, providing a significant boost to the search for clues leading to the accurate identification and effective treatment of cancers.
  17. 17
    Li, L.; Feng, J.; Liu, H.; Li, Q.; Tong, L.; Tang, B. Chem. Sci. 2016, 7, 1940– 1945 DOI: 10.1039/C5SC03909F
    Google Scholar
    17
    Two-color imaging of microRNA with enzyme-free signal amplification via hybridization chain reactions in living cells
    Li, Lu; Feng, Jie; Liu, Haiyun; Li, Qingling; Tong, Lili; Tang, Bo
    Chemical Science (2016), 7 (3), 1940-1945CODEN: CSHCCN; ISSN:2041-6520. (Royal Society of Chemistry)
    In situ imaging of miRNA in living cells could facilitate the monitoring of the dynamic expression and distribution of miRNA and research on miRNA-related cellular processes and diseases. Given the low expression levels and even down-regulation of cellular miRNA that is assocd. with some diseases, amplification strategies are imperative for intracellular miRNA imaging. The present paper proposes a non-destructive amplification strategy for use in living cells. This amplification strategy utilizes the enzyme-free hybridization chain reaction (HCR) with graphene oxide (GO) as a carrier to image cellular miRNA. The resulting signal amplification provides excellent recognition and signal enhancement of specific miRNAs in living cells. As the fluorescence quencher and probe carrier, GO enables activation of the signal switch and effective intracellular delivery of amplification reagents. This new imaging method realizes simple, sensitive and non-destructive signal amplification of miRNA in living cells and has an ability to simultaneously image two types of miRNA in the same cell. This method supplies accurate information regarding cellular miRNA-related biol. events and provides a new tool for highly sensitive and simultaneous imaging of multiple low-level biomarkers, thereby improving the accuracy of early disease diagnosis.
  18. 18
    Wu, Z.; Liu, G. Q.; Yang, X. L.; Jiang, J. J. Am. Chem. Soc. 2015, 137, 6829 DOI: 10.1021/jacs.5b01778
    Google Scholar
    18
    Electrostatic nucleic acid nanoassembly enables hybridization chain reaction in living cells for ultrasensitive mRNA imaging
    Wu, Zhan; Liu, Gao-Qin; Yang, Xiao-Li; Jiang, Jian-Hui
    Journal of the American Chemical Society (2015), 137 (21), 6829-6836CODEN: JACSAT; ISSN:0002-7863. (American Chemical Society)
    Efficient approaches for intracellular delivery of nucleic acid reagents to achieve sensitive detection and regulation of gene and protein expressions are essential for chem. and biol. We develop a novel electrostatic DNA nanoassembly that, for the first time, realizes hybridization chain reaction (HCR), a target-initiated alternating hybridization reaction between two hairpin probes, for signal amplification in living cells. The DNA nanoassembly has a designed structure with a core gold nanoparticle, a cationic peptide interlayer, and an electrostatically assembled outer layer of fluorophore-labeled hairpin DNA probes. It is shown to have high efficiency for cellular delivery of DNA probes via a unique endocytosis-independent mechanism that confers a significant advantage of overcoming endosomal entrapment. Moreover, electrostatic assembly of DNA probes enables target-initialized release of the probes from the nanoassembly via HCR. This intracellular HCR offers efficient signal amplification and enables ultrasensitive fluorescence activation imaging of mRNA expression with a picomolar detection limit. The results imply that the developed nanoassembly may provide an invaluable platform in low-abundance biomarker discovery and regulation for cell biol. and theranostics.
  19. 19
    Tang, Y.; Zhang, X.; Tang, L.; Yu, R.; Jiang, J. Anal. Chem. 2017, 89, 3445– 3451 DOI: 10.1021/acs.analchem.6b04312
    Google Scholar
    19
    In Situ Imaging of Individual mRNA Mutation in Single Cells Using Ligation-Mediated Branched Hybridization Chain Reaction (Ligation-bHCR)
    Tang, Ying; Zhang, Xiao-Li; Tang, Li-Juan; Yu, Ru-Qin; Jiang, Jian-Hui
    Analytical Chemistry (Washington, DC, United States) (2017), 89 (6), 3445-3451CODEN: ANCHAM; ISSN:0003-2700. (American Chemical Society)
    Ultrasensitive and specific in situ imaging of gene expression is essential for mol. medicine and clin. theranostics. The authors develop a novel fluorescence in situ hybridization (FISH) strategy based on a new branched hybridization chain reaction (bHCR) for efficient signal amplification in FISH assay and a ligase-mediated discrimination for specific mutation detection. To the knowledge, this is the first time that HCR has been realized for mutation detection in FISH assay. In vitro assay shows that the ligation-bHCR strategy affords high specificity in discriminating single-nucleotide variation in mRNA, and generates a highly branched polymeric product that confers more efficient amplification or better sensitivity than HCR. Imaging anal. reveals that ligation-bHCR generates highly bright spot-like signals for localization of individual mRNA mols., and spot signals of different colors are highly specific in genotyping point mutation of individual mRNA. Moreover, this strategy is shown to have the potential for quant. i.m.-aging of the expression of mRNA at the single cell level. Therefore, this strategy may provide a new promising paradigm in developing highly sensitive and specific FISH methods for various diagnostic and research applications.
  20. 20
    Xuan, F.; Hsing, I. M. J. Am. Chem. Soc. 2014, 136, 9810– 9813 DOI: 10.1021/ja502904s
    Google Scholar
    20
    Triggering hairpin-free chain-branching growth of fluorescent DNA dendrimers for nonlinear hybridization chain reaction
    Xuan, Feng; Hsing, I.-Ming
    Journal of the American Chemical Society (2014), 136 (28), 9810-9813CODEN: JACSAT; ISSN:0002-7863. (American Chemical Society)
    The authors present a nonlinear hybridization chain reaction (HCR) system in which a trigger DNA initiates self-sustained assembly of quenched double-stranded substrates into fluorescent dendritic nanostructures. During the process, an increasing no. of originally sequestered trigger sequences labeled with fluorescent reporters are freed up from quenched substrates, leading to chain-branching growth of the assembled DNA dendrimers and an exponential increase in the fluorescence intensity. The triggered assembly behavior was examd. by PAGE anal., and the morphologies of the grown dendrimers were verified by AFM imaging. The exponential kinetics of the fluorescence accumulation was also confirmed by time-dependent fluorescence spectroscopy. This method adopts a simple sequence design strategy, the concept of which could be adapted to program assembly systems with higher-order growth kinetics.
  21. 21
    Bi, S.; Chen, M.; Jia, X.; Dong, Y.; Wang, Z. Angew. Chem., Int. Ed. 2015, 54, 8144– 8148 DOI: 10.1002/anie.201501457
    Google Scholar
    There is no corresponding record for this reference.
  22. 22
    Chandran, H.; Rangnekar, A.; Shetty, G.; Schultes, E. A.; Reif, J. H.; LaBean, T. H. Biotechnol. J. 2013, 8, 221– 227 DOI: 10.1002/biot.201100499
    Google Scholar
    22
    An autonomously self-assembling dendritic DNA nanostructure for target DNA detection
    Chandran, Harish; Rangnekar, Abhijit; Shetty, Geetha; Schultes, Erik A.; Reif, John H.; LaBean, Thomas H.
    Biotechnology Journal (2013), 8 (2), 221-227CODEN: BJIOAM; ISSN:1860-6768. (Wiley-VCH Verlag GmbH & Co. KGaA)
    There is a growing need for sensitive and reliable nucleic acid detection methods that are convenient and inexpensive. Responsive and programmable DNA nanostructures have shown great promise as chem. detection systems. Here, we describe a DNA detection system employing the triggered self-assembly of a novel DNA dendritic nanostructure. The detection protocol is executed autonomously without external intervention. Detection begins when a specific, single-stranded target DNA strand (T) triggers a hybridization chain reaction (HCR) between two, distinct DNA hairpins (α and β). Each hairpin opens and hybridizes up to two copies of the other. In the absence of T, α and β are stable and remain in their poised, closed-hairpin form. In the presence of T, α hairpins are opened by toe-hold mediated strand-displacement, each of which then opens and hybridizes two β hairpins. Likewise, each opened β hairpin can open and hybridize two α hairpins. Hence, each layer of the growing dendritic nanostructure can in principle accommodate an exponentially increasing no. of cognate mols., generating a high mol. wt. nanostructure. This HCR system has minimal sequence constraints, allowing reconfiguration for the detection of arbitrary target sequences. Here, we demonstrate detection of unique sequence identifiers of HIV and Chlamydia pathogens.
  23. 23
    Wei, J.; Gong, X.; Wang, Q.; Pan, M.; Liu, X.; Liu, J.; Xia, F.; Wang, F. Chem. Sci. 2018, 9, 52 DOI: 10.1039/C7SC03939E
    Google Scholar
    23
    Construction of an autonomously concatenated hybridization chain reaction for signal amplification and intracellular imaging
    Wei, Jie; Gong, Xue; Wang, Qing; Pan, Min; Liu, Xiaoqing; Liu, Jing; Xia, Fan; Wang, Fuan
    Chemical Science (2018), 9 (1), 52-61CODEN: CSHCCN; ISSN:2041-6520. (Royal Society of Chemistry)
    Biomol. self-assembly has spurred substantial research efforts for the development of low-cost point-of-care diagnostics. Herein, we introduce an isothermal enzyme-free concatenated hybridization chain reaction (C-HCR), in which the output of the upstream hybridization chain reaction (HCR-1) layer acts as an intermediate input to activate the downstream hybridization chain reaction (HCR-2) layer. The initiator motivates HCR-1 through the autonomous cross-opening of two functional DNA hairpins, yielding polymeric dsDNA nanowires composed of numerous tandem triggers T as output of the primary sensing event. The reconstituted amplicon T then initiates HCR-2 and transduces the analyte recognition into an amplified readout, originating from the synergistic effect between HCR-1 and HCR-2 layers. Native gel electrophoresis, atom force microscopy (AFM) and fluorescence spectra revealed that C-HCR mediated the formation of frond-like branched dsDNA nanowires and the generation of an amplified FRET signal. As a versatile and robust amplification strategy, the unpreceded C-HCR can discriminate DNA analyte from its mutants with high accuracy and specificity. By incorporating an auxiliary sensing module, the integrated C-HCR amplifier was further adapted for highly sensitive and selective detection of microRNA (miRNA), as a result of the hierarchical and sequential hybridization chain reactions, in human serum and even living cells through an easy-to-integrate "plug-and-play" procedure. In addn., the C-HCR amplifier was successfully implemented for intracellular miRNA imaging by acquiring an accurate and precise signal localization inside living cells, which was esp. suitable for the ex situ and in situ amplified detection of trace amts. of analyte. The C-HCR amplification provides a comprehensive and smart toolbox for highly sensitive detection of various biomarkers and thus should hold great promise in clin. diagnosis and assessment. The infinite layer of multilayered C-HCR is anticipated to further strengthen the amplification capacity and reliability (anti-invasion performance) of intracellular imaging approach, which is of great significance for its bioanal. applications.
  24. 24
    Chi, Y.; Wang, X.; Yang, Y.; Zhang, C.; Ertl, H. C.; Zhou, D. Mol. Ther.--Nucleic Acids 2014, 3, e208 DOI: 10.1038/mtna.2014.59
    Google Scholar
    24
    Survivin-targeting Artificial MicroRNAs Mediated by Adenovirus Suppress Tumor Activity in Cancer Cells and Xenograft Models
    Chi, Yudan; Wang, Xiang; Yang, Yong; Zhang, Chao; Ertl, Hildegund C. J.; Zhou, Dongming
    Molecular Therapy--Nucleic Acids (2014), 3 (11), e208CODEN: MTAOC5; ISSN:2162-2531. (Nature Publishing Group)
    Survivin is highly expressed in most human tumors and fetal tissue, and absent in terminally differentiated cells. It promotes tumor cell proliferation by neg. regulating cell apoptosis and facilitating cell division. Survivin's selective expression pattern suggests that it might be a suitable target for cancer therapy, which would promote death of transformed but not normal cells. This was tested using artificial microRNAs (amiRNAs) targeting survivin. After screening, two effective amiRNAs, which knocked down survivin expression, were identified and cloned into a replication-defective adenoviral vector. Tumor cells infected with the recombinant vector downregulated expression of survivin and underwent apoptotic cell death. Further studies showed that apoptosis was assocd. with increases in caspase 3 and cleaved Poly (ADP-ribose) polymerase, and activation of the p53 signaling pathway. Furthermore, amiRNA treatment caused blockade of mitosis and cell cycle arrest at the G2/M phase. In vivo, survivin-targeting amiRNAs expressed by adenoviral vectors effectively delayed growth of hepatocellular and cervical carcinomas in mouse xenograft models. These results indicate that silencing of survivin by amiRNA has potential for treatment of cancer.
  25. 25
    Peng, X. H.; Cao, Z. H.; Xia, J. T.; Carlson, G. W.; Lewis, M. M.; Wood, W. C.; Yang, L. Cancer Res. 2005, 65, 1909– 1917 DOI: 10.1158/0008-5472.CAN-04-3196
    Google Scholar
    There is no corresponding record for this reference.
  26. 26
    Yamanaka, K.; Nakahara, T.; Yamauchi, T.; Kita, A.; Takeuchi, M.; Kiyonaga, F.; Kaneko, N.; Sasamata, M. Clin. Cancer Res. 2011, 17, 5423– 31 DOI: 10.1158/1078-0432.CCR-10-3410
    Google Scholar
    26
    Antitumor Activity of YM155, a Selective Small-Molecule Survivin Suppressant, Alone and in Combination with Docetaxel in Human Malignant Melanoma Models
    Yamanaka, Kentaro; Nakahara, Takahito; Yamauchi, Tomohiro; Kita, Aya; Takeuchi, Masahiro; Kiyonaga, Fumiko; Kaneko, Naoki; Sasamata, Masao
    Clinical Cancer Research (2011), 17 (16), 5423-5431CODEN: CCREF4; ISSN:1078-0432. (American Association for Cancer Research)
    Purpose: Aggressive cell growth and chemoresistance are notorious obstacles in melanoma therapy. Accumulating evidence suggests that survivin is preferentially expressed in cancer cells and plays a crucial role in cell division and apoptosis dysfunction. Here, we evaluated the therapeutic potential of YM155, a selective survivin suppressant, alone and in combination with docetaxel using human melanoma models. Exptl. Design: A375 and SK-MEL-5 human malignant melanoma cells were treated with siRNA, YM155, and/or docetaxel, and cell viability, mRNA and protein expression levels, cell-cycle distribution, and immunohistochem. staining were then evaluated. Furthermore, the efficacy of YM155 combined with docetaxel was further examd. in established xenograft models. Results: Survivin suppression was sufficient to induce spontaneous apoptosis of melanoma cells. YM155 showed nanomolar antiproliferative effects and induced tumor regression in established melanoma xenograft models. Docetaxel showed antitumor activity against melanoma cells, although it also induced survivin upregulation and G2/M mitotic arrest; however, cotreatment with YM155 decreased survivin expression below basal levels. Combination treatment of YM155 and docetaxel induced a greater rate of apoptosis than the sum of the single-treatment rates and promoted tumor regression without enhanced body wt. loss in the melanoma xenograft models. Conclusions: Survivin is responsible for the inherent low levels of spontaneous apoptosis in melanoma cells. The concomitant combination of YM155 with docetaxel diminished the accumulation of survivin in G2/M mitotic arrest, and induced more intense apoptosis compared with each single treatment. YM155 in combination with docetaxel is well tolerated and shows greater efficacy than either agent alone in mouse xenograft models. Clin Cancer Res; 17(16); 5423-31.

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  2. Mengdie Li, Jiayang He, Xin Shang, Chunli Yang, Yuqing Zhang, Siyu Zuo, Ruo Yuan, Wenju Xu. A Reciprocal-Amplifiable Fluorescence Sensing Platform via Replicated Hybridization Chain Reaction for Hosting Concatenated Multi-Ag Nanoclusters as Signal Reporter. Analytical Chemistry 2022, 94 (47) , 16427-16435. https://doi.org/10.1021/acs.analchem.2c03782
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  4. Dan-Dan Wang, Juan Zhang, Qiao-Qin Yu, Ke Zhang, Ting-Ting Chen, Xia Chu. Biomineralized Zeolitic Imidazolate Framework-8 Nanoparticles Enable Polymerase-Driven DNA Biocomputing for Reliable Cell Identification. Analytical Chemistry 2022, 94 (11) , 4794-4802. https://doi.org/10.1021/acs.analchem.1c05605
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  7. Hua Chai, Wenbo Cheng, Dayong Jin, Peng Miao. Recent Progress in DNA Hybridization Chain Reaction Strategies for Amplified Biosensing. ACS Applied Materials & Interfaces 2021, 13 (33) , 38931-38946. https://doi.org/10.1021/acsami.1c09000
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  11. Xueli Zhu, Bin Qu, Zhan-Ming Ying, Jin-Wen Liu, Zhenkun Wu, Ru-Qin Yu, Jian-Hui Jiang. Cascade Circuits on Self-Assembled DNA Polymers for Targeted RNA Imaging In Vivo. Analytical Chemistry 2020, 92 (24) , 15953-15958. https://doi.org/10.1021/acs.analchem.0c03400
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  19. Chengcheng Gu, Xinke Kong, Xiaojuan Liu, Panpan Gai, Feng Li. Enzymatic Biofuel-Cell-Based Self-Powered Biosensor Integrated with DNA Amplification Strategy for Ultrasensitive Detection of Single-Nucleotide Polymorphism. Analytical Chemistry 2019, 91 (13) , 8697-8704. https://doi.org/10.1021/acs.analchem.9b02510
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  22. Yanxin Chu, An-Ping Deng, Wenjing Wang, Jun-Jie Zhu. Concatenated Catalytic Hairpin Assembly/Hyperbranched Hybridization Chain Reaction Based Enzyme-Free Signal Amplification for the Sensitive Photoelectrochemical Detection of Human Telomerase RNA. Analytical Chemistry 2019, 91 (5) , 3619-3627. https://doi.org/10.1021/acs.analchem.8b05610
  23. Hongfei He, Jianyuan Dai, Guixiu Dong, Hongli Shi, Fang Wang, Yunran Qiu, Ruoxing Liao, Cuisong Zhou, Yong Guo, Dan Xiao. Self-Replication-Assisted Rapid Preparation of DNA Nanowires at Room Temperature and Its Biosensing Application. Analytical Chemistry 2019, 91 (4) , 3043-3047. https://doi.org/10.1021/acs.analchem.8b05431
  24. Lei Yang, Qiong Wu, Yuqi Chen, Xiaoqing Liu, Fuan Wang, Xiang Zhou. Amplified MicroRNA Detection and Intracellular Imaging Based on an Autonomous and Catalytic Assembly of DNAzyme. ACS Sensors 2019, 4 (1) , 110-117. https://doi.org/10.1021/acssensors.8b01000
  25. Xudong Wang, Jun Dai, Xuehong Min, Zhihua Yu, Yong Cheng, Kaixun Huang, Juliang Yang, Xiaoqing Yi, Xiaoding Lou, Fan Xia. DNA-Conjugated Amphiphilic Aggregation-Induced Emission Probe for Cancer Tissue Imaging and Prognosis Analysis. Analytical Chemistry 2018, 90 (13) , 8162-8169. https://doi.org/10.1021/acs.analchem.8b01456
  26. Sayantan Tripathy, Ng Ka Wai, Sahil Sharma, Ngoc Nhu Vu, Siddhant Jaitpal, Fatemah Zare, Muskaan Singh, Liam McDaid, Saugat Bhattacharyya, Gerard L. Coté, Samuel B. Mabbott. A Locked Aptamer-Magnetic Nanoparticle Assay for Cardiac Troponin I Classification to Support Myocardial Infarction Diagnosis in Resource-Limited Environments. Biosensors and Bioelectronics 2025, 32 , 117590. https://doi.org/10.1016/j.bios.2025.117590
  27. Jinglin Fu, Qiaochu Zhang, Shiming Liu, Derek Puyat, Akshay Shah, Alireza Ebrahimimojarad, Sung Won Oh. Rapid Amplification and Detection of Single‐Stranded Nucleic Acids for Point‐of‐Care Diagnosis. Small Methods 2025, 37 https://doi.org/10.1002/smtd.202401733
  28. Su-Jing Zhao, Zhenkun Wu, Jian-Hui Jiang. A Fluorogenic Sensor via Catalytic Hairpin Assembly for Precise Live-Cell Imaging of mRNA. 2025, 165-175. https://doi.org/10.1007/978-1-0716-4248-1_14
  29. Ting Chen, Xiaojiao Wang, Rongrong Gao, Meihe Yuan, Mei Chen, Xiao-Bing Zhang, Guoliang Ke. Size-selective hybridization chain reaction for accurate signal amplification in living cancer cells. Science China Chemistry 2024, 67 (12) , 4259-4266. https://doi.org/10.1007/s11426-024-2294-x
  30. Hang Gong, Shufen Yao, Xiaojia Zhao, Feng Chen, Chunyan Chen, Changqun Cai. DNA nanosensor based on bipedal 3D DNA walker–driven proximal catalytic hairpin assembly for sensitive and fast TK1 mRNA detection. Microchimica Acta 2024, 191 (8) https://doi.org/10.1007/s00604-024-06569-w
  31. Mingzhu Fan, Xiaohong Zhong, Yanshi Wei, Rong Wang, Yifang He, Yuhui Liu, Xin Huang, Zhihui Luo, Jiayao Xu, Shulong Wang, Yong Huang. A smart mRNA-initiated DNAzyme nanogenerator for precise imaging and gene therapy in vivo. Sensors and Actuators B: Chemical 2024, 411 , 135742. https://doi.org/10.1016/j.snb.2024.135742
  32. Xing Lu, Xujiao Zhou, Bin Song, Hong Zhang, Mingrui Cheng, Xingyu Zhu, Yuqi Wu, Haoliang Shi, Binbin Chu, Yao He, Houyu Wang, Jiaxu Hong. Framework Nucleic Acids Combined with 3D Hybridization Chain Reaction Amplifiers for Monitoring Multiple Human Tear Cytokines. Advanced Materials 2024, 36 (26) https://doi.org/10.1002/adma.202400622
  33. Qiuyan Huang, Kun Wang, Yuan Wang. Highly sensitive miRNA-21 detection with enzyme-free cascade amplification biosensor. Talanta 2024, 273 , 125928. https://doi.org/10.1016/j.talanta.2024.125928
  34. Xiaojing Xing, Mengying Gao, Minglin Lei, Kunqi Cheng, Yifan Zhao, Xianchao Du, Luyi Zong, Dongfang Qiu, Xueguo Liu. MOF-mediated dual energy transfer nanoprobe integrated with exonuclease III amplification strategy for highly sensitive detection of DNA. Analytical Methods 2024, 16 (13) , 1916-1922. https://doi.org/10.1039/D4AY00127C
  35. Cong Fan, Longjie Xie, Feng Zhao, Jingjing Wang, Xiandong Lin, Xian Chen. Novel fluorescence nano-orbital biosensor for highly sensitive microRNA detection. Analytica Chimica Acta 2024, 1288 , 342172. https://doi.org/10.1016/j.aca.2023.342172
  36. Yousuke Tsuneoka, Hiromasa Funato. Whole Brain Mapping of Orexin Receptor mRNA Expression Visualized by Branched In Situ Hybridization Chain Reaction. eneuro 2024, 11 (2) , ENEURO.0474-23.2024. https://doi.org/10.1523/ENEURO.0474-23.2024
  37. Zhenli Qiu, Xintong Lin, Yufen Lei, Jinman Zhu, Rongjian Sa, Yiting Chen. Contactless photoelectrochemical biosensors based on hierarchical MXene/Bi2S3 nanosheets with the branched hybridization chain reaction. Biosensors and Bioelectronics 2024, 243 , 115764. https://doi.org/10.1016/j.bios.2023.115764
  38. Yuanyuan Chen, Hui Jiang, Xiaohui Liu, Xuemei Wang. Engineered Electrochemiluminescence Biosensors for Monitoring Heavy Metal Ions: Current Status and Prospects. Biosensors 2024, 14 (1) , 9. https://doi.org/10.3390/bios14010009
  39. Lie Li, Suping Li, Jie Wang, Xiaohong Wen, Mei Yang, Haiyan Chen, Qiuping Guo, Kemin Wang. Extracellular ATP-activated hybridization chain reaction for accurate and sensitive detection of cancer cells. Chinese Chemical Letters 2023, 34 (12) , 108399. https://doi.org/10.1016/j.cclet.2023.108399
  40. Wenjia Wang, Qinyu Ge, Xiangwei Zhao. Enzyme-free isothermal amplification strategy for the detection of tumor-associated biomarkers: A review. TrAC Trends in Analytical Chemistry 2023, 160 , 116960. https://doi.org/10.1016/j.trac.2023.116960
  41. Chunhuan Xu, Jingjin Zhao, Shengyu Chen, Ivan Yu Sakharov, Shengqiang Hu, Shulin Zhao. An ultrasensitive bunge bedstraw herb type DNA machine for absolute quantification of mRNA in single cell. Biosensors and Bioelectronics 2022, 217 , 114702. https://doi.org/10.1016/j.bios.2022.114702
  42. Xianghui Li, Yichan Huang, Jiawen Chen, Shuangmu Zhuo, Zhenyu Lin, Jianxin Chen. A highly sensitive homogeneous electrochemiluminescence biosensor for flap endonuclease 1 based on branched hybridization chain reaction amplification and ultrafiltration separation. Bioelectrochemistry 2022, 147 , 108189. https://doi.org/10.1016/j.bioelechem.2022.108189
  43. Xiaowei Wei, Zehan Cai, Lijun Cai, Yu Wang, Dagan Zhang, Yefei Zhu. Photonic Barcodes Combining Branched Hybridization Chain Reaction for Multiplex Quantification of Bladder Cancer MicroRNAs. Advanced Materials Interfaces 2022, 9 (20) https://doi.org/10.1002/admi.202102515
  44. Sima Khajouei, Elyas Hosseinzadeh, Hadi Ravan, Abbas Mohammadi. Binary detection of protein and nucleic acid enabled cancer diagnosis through branched hybridization chain reaction. Analytica Chimica Acta 2022, 1205 , 339755. https://doi.org/10.1016/j.aca.2022.339755
  45. Yuning Wang, Bin Li, Tongtong Tian, Yujie Liu, Juxiang Zhang, Kun Qian. Advanced on-site and in vitro signal amplification biosensors for biomolecule analysis. TrAC Trends in Analytical Chemistry 2022, 149 , 116565. https://doi.org/10.1016/j.trac.2022.116565
  46. Shiyuan Liu, Zhenxiang Liu, Huimin Wang, Jiaoli Wang, Ruiting Liu, Kemin Wang, Jin Huang. A novel FRET-based dendritic hybridization chain reaction for tumour-related mRNA imaging. Chemical Communications 2022, 58 (9) , 1414-1417. https://doi.org/10.1039/D1CC06009K
  47. Tingting Xu, Yao Sun, Sha Yu, Shaojun Wu, Yu Su, Ye Tian, Yuanzhen Zhou, Jun-Jie Zhu. A fluorogenic RNA aptamer nanodevice for the low background imaging of mRNA in living cells. Chemical Communications 2022, 58 (9) , 1354-1357. https://doi.org/10.1039/D1CC06582C
  48. Shulin Zhao, Chunhuan Xu, Jingjin Zhao, Shengyu Chen, Ivan Sakharov, Shengqiang Hu. An Ultrasensitive Bunge Bedstraw Herb Type DNA Machine for Absolute Quantification of Mrna in Single Cell. SSRN Electronic Journal 2022, 3 https://doi.org/10.2139/ssrn.4183372
  49. Jingting Wu, Jinrui Lv, Xiaoqi Zheng, Zai-Sheng Wu. Hybridization chain reaction and its applications in biosensing. Talanta 2021, 234 , 122637. https://doi.org/10.1016/j.talanta.2021.122637
  50. Su Li, Yuyin Tang, Linyu Hu, Wenhui Qian, Lijuan Ma, Xuetong Sun, Wenwen Ji, Bei Zhang, Shuying Han, Yumei Chi, Dong Zhu. Autonomously driving multiplexed hierarchical hybridization chain reaction of a DNA cobweb sensor for monitoring intracellular microRNA. Sensors and Actuators B: Chemical 2021, 344 , 130205. https://doi.org/10.1016/j.snb.2021.130205
  51. Zackary A. Zimmers, Alexander D. Boyd, Hannah E. Stepp, Nicholas M. Adams, Frederick R. Haselton. Development of an Automated, Non-Enzymatic Nucleic Acid Amplification Test. Micromachines 2021, 12 (10) , 1204. https://doi.org/10.3390/mi12101204
  52. Long Jiang, Xiaonan Qu, Wenyu Sun, Manru Zhang, Yeru Wang, Yu Wang, Yijin Zhao, Fenfen Zhang, Yuning Leng, Su Liu, Jinghua Yu, Jiadong Huang. A three-dimensional dynamic DNA walker-mediated branching hybridization chain reaction for the ultrasensitive fluorescence sensing of ampicillin. The Analyst 2021, 146 (17) , 5413-5420. https://doi.org/10.1039/D0AN02226H
  53. Xiu Liu, Shengjun Bu, Hongguo Wei, Ze Wang, Songling Yu, Zhongyi Li, Zhuo Hao, Xiuxia He, Jiayu Wan. Visual assay of Escherichia coli O157:H7 based on an isothermal strand displacement and hybrid chain reaction amplification strategy. Analytical Methods 2021, 13 (30) , 3379-3385. https://doi.org/10.1039/D1AY00644D
  54. Meng-Qi He, Shuai Chen, Wei Shi, Xue-Wei Zhang, Yong-Liang Yu, Jian-Hua Wang. All-in-one fractal nanoplasmonic array for visual discrimination of biomolecules and microorganisms. Sensors and Actuators B: Chemical 2021, 338 , 129832. https://doi.org/10.1016/j.snb.2021.129832
  55. Yuqi Huang, Shuhui Zhao, Wenxiu Zhang, Qiuyue Duan, Qi Yan, Hu Fu, Liang Zhong, Gang Yi. Multifunctional electrochemical biosensor with “tetrahedral tripods” assisted multiple tandem hairpins assembly for ultra-sensitive detection of target DNA. RSC Advances 2021, 11 (33) , 20046-20056. https://doi.org/10.1039/D1RA02424H
  56. Idorenyin A. Iwe, Wenqin Li, Zhigang Li, Jiahao Huang. Hairpin DNA-Mediated isothermal amplification (HDMIA) techniques for nucleic acid testing. Talanta 2021, 226 , 122146. https://doi.org/10.1016/j.talanta.2021.122146
  57. Jiafeng Pan, Ying He, Zhi Liu, Junhua Chen. Dual recognition element-controlled logic DNA circuit for COVID-19 detection based on exonuclease III and DNAzyme. Chemical Communications 2021, 57 (9) , 1125-1128. https://doi.org/10.1039/D0CC06799G
  58. Na Wu, Yi-Ting Wang, Xiao-Yan Wang, Xu- Wei Chen, Ting Yang, Jian-Hua Wang. A simple, one-pot and ultrasensitive DNA sensor via Exo III-Assisted target recycling and 3D DNA walker cascade amplification. Analytica Chimica Acta 2021, 1147 , 15-22. https://doi.org/10.1016/j.aca.2020.12.026
  59. Zhuoer Zeng, Rong Zhou, Ruowei Sun, Xun Zhang, Zeneng Cheng, Chuanpin Chen, Qubo Zhu. Nonlinear hybridization chain reaction-based functional DNA nanostructure assembly for biosensing, bioimaging applications. Biosensors and Bioelectronics 2021, 173 , 112814. https://doi.org/10.1016/j.bios.2020.112814
  60. Xiaohao Liu, Sinuo Yu, Chang Feng, Dongsheng Mao, Jinlong Li, Xiaoli Zhu. In situ Analysis of Cancer Cells Based on DNA Signal Amplification and DNA Nanodevices. Critical Reviews in Analytical Chemistry 2021, 51 (1) , 8-19. https://doi.org/10.1080/10408347.2019.1674631
  61. Na Li, Fuzhi Shen, Zhiheng Cai, Wenzhen Pan, Yiming Yin, Xuan Deng, Xing Zhang, Jeremiah Ong'achwa Machuki, Yanyan Yu, Dongzhi Yang, Yun Yang, Ming Guan, Fenglei Gao. Target‐Induced Core–Satellite Nanostructure Assembly Strategy for Dual‐Signal‐On Fluorescence Imaging and Raman Quantification of Intracellular MicroRNA Guided Photothermal Therapy. Small 2020, 16 (49) https://doi.org/10.1002/smll.202005511
  62. Yan Zhao, Yuanbo Feng, Yuanbo Zhang, Pu Xia, Zihan Xiao, Ziheng Wang, Hongxia Yan. Combining competitive sequestration with nonlinear hybridization chain reaction amplification: an ultra-specific and highly sensitive sensing strategy for single-nucleotide variants. Analytica Chimica Acta 2020, 1130 , 107-116. https://doi.org/10.1016/j.aca.2020.07.022
  63. Shiyan Dai, Yuting Zhou, Guifang Cheng, Pingang He, Yuzhi Fang. Dual-signal electrochemical sensor for detection of cancer cells by the split primer ligation-triggered catalyzed hairpin assembly. Talanta 2020, 217 , 121079. https://doi.org/10.1016/j.talanta.2020.121079
  64. Elyas Hosseinzadeh, Hadi Ravan, Abbas Mohammadi, Hossein Pourghadamyari. Colorimetric detection of miRNA-21 by DNAzyme-coupled branched DNA constructs. Talanta 2020, 216 , 120913. https://doi.org/10.1016/j.talanta.2020.120913
  65. Juan Zhang, Mengyun He, Cunpeng Nie, Manman He, Qingshan Pan, Chang Liu, Yanlei Hu, Tingting Chen, Xia Chu. Biomineralized metal–organic framework nanoparticles enable a primer exchange reaction-based DNA machine to work in living cells for imaging and gene therapy. Chemical Science 2020, 11 (27) , 7092-7101. https://doi.org/10.1039/D0SC00339E
  66. Yousuke Tsuneoka, Hiromasa Funato. Modified in situ Hybridization Chain Reaction Using Short Hairpin DNAs. Frontiers in Molecular Neuroscience 2020, 13 https://doi.org/10.3389/fnmol.2020.00075
  67. Devleena Samanta, Sasha B. Ebrahimi, Chad A. Mirkin. Nucleic‐Acid Structures as Intracellular Probes for Live Cells. Advanced Materials 2020, 32 (13) https://doi.org/10.1002/adma.201901743
  68. Kaixiang Zhang, Ruijie Deng, Hua Gao, Xucong Teng, Jinghong Li. Lighting up single-nucleotide variation in situ in single cells and tissues. Chemical Society Reviews 2020, 49 (6) , 1932-1954. https://doi.org/10.1039/C9CS00438F
  69. Guang-Li Li, Yong-Hui Xia, Qing Li. Biodegradable MnO2 nanosheet-mediated catalytic hairpin assembly for two-color imaging of mRNAs in living cells. Microchemical Journal 2020, 153 , 104367. https://doi.org/10.1016/j.microc.2019.104367
  70. Ke Quan, Cuiping Yi, Xiaohai Yang, Xiaoxiao He, Jin Huang, Kemin Wang. FRET-based nucleic acid probes: Basic designs and applications in bioimaging. TrAC Trends in Analytical Chemistry 2020, 124 , 115784. https://doi.org/10.1016/j.trac.2019.115784
  71. Manman He, Mengyun He, Juan Zhang, Chang Liu, Qingshan Pan, Jintao Yi, Tingting Chen. A spatial-confinement hairpin cascade reaction-based DNA tetrahedral amplifier for mRNA imaging in live cells. Talanta 2020, 207 , 120287. https://doi.org/10.1016/j.talanta.2019.120287
  72. Dan Zhao, Qingxin Yin, Yangyang Chang, Meng Liu. Nucleic acid circuits for cell imaging: From the test tube to the cell. TrAC Trends in Analytical Chemistry 2020, 122 , 115706. https://doi.org/10.1016/j.trac.2019.115706
  73. Gaolian Xu, Mingliang Lai, Rab Wilson, Andrew Glidle, Julien Reboud, Jonathan M. Cooper. Branched hybridization chain reaction—using highly dimensional DNA nanostructures for label-free, reagent-less, multiplexed molecular diagnostics. Microsystems & Nanoengineering 2019, 5 (1) https://doi.org/10.1038/s41378-019-0076-z
  74. Yao Qin, Daxiu Li, Ruo Yuan, Yun Xiang. Netlike hybridization chain reaction assembly of DNA nanostructures enables exceptional signal amplification for sensing trace cytokines. Nanoscale 2019, 11 (35) , 16362-16367. https://doi.org/10.1039/C9NR04988F
  75. Kumar Vikrant, Neha Bhardwaj, Sanjeev K. Bhardwaj, Ki-Hyun Kim, Akash Deep. Nanomaterials as efficient platforms for sensing DNA. Biomaterials 2019, 214 , 119215. https://doi.org/10.1016/j.biomaterials.2019.05.026
  76. Zhihe Qing, Jingyuan Xu, Jinlei Hu, Jing Zheng, Lei He, Zhen Zou, Sheng Yang, Weihong Tan, Ronghua Yang. In Situ Amplification‐Based Imaging of RNA in Living Cells. Angewandte Chemie 2019, 131 (34) , 11698-11709. https://doi.org/10.1002/ange.201812449
  77. Zhihe Qing, Jingyuan Xu, Jinlei Hu, Jing Zheng, Lei He, Zhen Zou, Sheng Yang, Weihong Tan, Ronghua Yang. In Situ Amplification‐Based Imaging of RNA in Living Cells. Angewandte Chemie International Edition 2019, 58 (34) , 11574-11585. https://doi.org/10.1002/anie.201812449
  78. Takashi Sakamoto, Rikuto Yamada. Label‐Free in Situ Monitoring of the DNA Hybridization Chain Reaction by Using Sequence‐Selective Minor‐Groove‐Binding Fluorophores. ChemBioChem 2019, 20 (10) , 1242-1245. https://doi.org/10.1002/cbic.201800738
  79. Ruijin Zeng, Lijia Zhang, Lingshan Su, Zhongbin Luo, Qian Zhou, Dianping Tang. Photoelectrochemical bioanalysis of antibiotics on rGO-Bi2WO6-Au based on branched hybridization chain reaction. Biosensors and Bioelectronics 2019, 133 , 100-106. https://doi.org/10.1016/j.bios.2019.02.067
  80. Du-Juan Huang, Ting Cao, Zhi-Mei Huang, Zhenkun Wu, Li-Juan Tang, Jian-Hui Jiang. Crosslinking catalytic hairpin assembly for high-contrast imaging of multiple mRNAs in living cells. Chemical Communications 2019, 55 (27) , 3899-3902. https://doi.org/10.1039/C9CC01033E
  81. Kyeonghye Guk, Seul Gee Hwang, Jaewoo Lim, Hye-young Son, Yuna Choi, Yong-Min Huh, Taejoon Kang, Juyeon Jung, Eun-Kyung Lim. Fluorescence amplified sensing platforms enabling miRNA detection by self-circulation of a molecular beacon circuit. Chemical Communications 2019, 55 (24) , 3457-3460. https://doi.org/10.1039/C9CC00351G
  82. Jun Yang, Ben Q. Li, Rongheng Li, Xuesong Mei. Quantum 3D thermal imaging at the micro–nanoscale. Nanoscale 2019, 11 (5) , 2249-2263. https://doi.org/10.1039/C8NR09096C
  83. Qiong Li. Nucleic Acid Amplification Strategy-Based Fluorescence Imaging. 2019, 213-240. https://doi.org/10.1007/978-981-13-7044-1_11
  84. Dinggeng He, Ka-Wang Wong, Zhenzhen Dong, Hung-Wing Li. Recent progress in live cell mRNA/microRNA imaging probes based on smart and versatile nanomaterials. Journal of Materials Chemistry B 2018, 6 (47) , 7773-7793. https://doi.org/10.1039/C8TB02285B
  85. Peng Zhang, Jennifer Crow, Divya Lella, Xin Zhou, Glenson Samuel, Andrew K. Godwin, Yong Zeng. Ultrasensitive quantification of tumor mRNAs in extracellular vesicles with an integrated microfluidic digital analysis chip. Lab on a Chip 2018, 18 (24) , 3790-3801. https://doi.org/10.1039/C8LC01071D
  86. Xiyue Xie, Yaqin Chai, Yali Yuan, Ruo Yuan. Dual triggers induced disassembly of DNA polymer decorated silver nanoparticle for ultrasensitive electrochemical Pb2+ detection. Analytica Chimica Acta 2018, 1034 , 56-62. https://doi.org/10.1016/j.aca.2018.06.050
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  • Abstract

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    Scheme 1

    Scheme 1. Working Principle of bHCR for mRNA Imaging
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    Figure 1

    Figure 1. (a) Gel electrophoresis analysis of bHCR: lane 1, 500 nM H1 and 500 nM H2 with 250 nM T; lane 2, 1 μM H3 and 1 μM H4 with 500 nM T2; lane 3, 500 nM H1, 500 nM H2, 1 μM H3 with 250 nM T; lane 4, 500 nM H1, 500 nM H2, 1 μM H3 and 1 μM H4 with 250 nM T; lane 5, 500 nM H1, 500 nM H2, 1 μM H3 and 1 μM H4. (b–f) AFM images of the assembly products in primary HCR circuit (b), secondary HCR circuit (c), bHCR system in the presence of T (d), and absence of T (e). Part f is enlarged details of part d. Scale bar: 200 nm.

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    Figure 2

    Figure 2. (a) Fluorescence spectral responses for mixtures of 200 nM H1 and 200 nM H2 (blue); 200 nM F–H1 and 200 nM H2 with 2 nM T (green); 200 nM H1 and 200 nM H2, 200 nM F–H3 and 200 nM H4 (black); 200 nM H1, 200 nM H2, 200 nM F–H3, and 200 nM H4 with 2 nM T (pink); or with 2 nM one-base mismatched T (Mu-T) (red). (b) Fluorescence responses of bHCR system to T of varying concentrations. (c) Calibration curves of fluorescence intensities versus target concentrations. Inset shows the linear relationship between fluorescence intensities and target concentrations in the linear range. (d) Histograms of signal-to-background (STB) ratios of bHCR system (blue) and linear system (red) at different target concentrations.

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    Figure 3

    Figure 3. Fluorescence images for (a) HeLa cells transfected with H1, H2, F–H3, and H4; (b) C166 cells transfected with H1, H2, and F–H3 H4; (c) HeLa cells transfected with F–H1 and H2. (1) Fluorescence; (2) merged with DIC. Scale bar: 20 μm.

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  • References

    This article references 26 other publications.

    1. 1
      Tyagi, S. Nat. Methods 2009, 6, 331– 338 DOI: 10.1038/nmeth.1321
      1
      Imaging intracellular RNA distribution and dynamics in living cells
      Tyagi, Sanjay
      Nature Methods (2009), 6 (5), 331-338CODEN: NMAEA3; ISSN:1548-7091. (Nature Publishing Group)
      A review. Powerful methods now allow the imaging of specific mRNAs in living cells. These methods enlist fluorescent proteins to illuminate mRNAs, use labeled oligonucleotide probes and exploit aptamers that render org. dyes fluorescent. The intracellular dynamics of mRNA synthesis, transport and localization can be analyzed at higher temporal resoln. with these methods than has been possible with traditional fixed-cell or biochem. approaches. These methods have also been adopted to visualize and track single mRNA mols. in real time. This review explores the promises and limitations of these methods.
    2. 2
      Xia, Y.; Zhang, R.; Wang, Z.; Tian, J.; Chen, X. Chem. Soc. Rev. 2017, 46, 2824– 2843 DOI: 10.1039/C6CS00675B
      2
      Recent advances in high-performance fluorescent and bioluminescent RNA imaging probes
      Xia, Yuqiong; Zhang, Ruili; Wang, Zhongliang; Tian, Jie; Chen, Xiaoyuan
      Chemical Society Reviews (2017), 46 (10), 2824-2843CODEN: CSRVBR; ISSN:0306-0012. (Royal Society of Chemistry)
      RNA plays an important role in life processes. Imaging of mRNAs (mRNAs) and micro-RNAs (miRNAs) not only allows us to learn the formation and transcription of mRNAs and the biogenesis of miRNAs involved in various life processes, but also helps in detecting cancer. High-performance RNA imaging probes greatly expand our view of life processes and enhance the cancer detection accuracy. In this review, we summarize the state-of-the-art high-performance RNA imaging probes, including exogenous probes that can image RNA sequences with special modification and endogeneous probes that can directly image endogenous RNAs without special treatment. For each probe, we review its structure and imaging principle in detail. Finally, we summarize the application of mRNA and miRNA imaging probes in studying life processes as well as in detecting cancer. By correlating the structures and principles of various probes with their practical uses, we compare different RNA imaging probes and offer guidance for better utilization of the current imaging probes and the future design of higher-performance RNA imaging probes.
    3. 3
      Tyagi, S.; Kramer, F. R. Nat. Biotechnol. 1996, 14, 303– 308 DOI: 10.1038/nbt0396-303
      3
      Molecular beacons: probes that fluoresce upon hybridization
      Tyagi, Sanjay; Kramer, Fred Russell
      Nature Biotechnology (1996), 14 (3), 303-8CODEN: NABIF9; ISSN:1087-0156. (Nature Publishing Co.)
      The authors have developed novel nucleic acid probes that recognize and report the presence of specific nucleic acids in homogeneous solns. These probes undergo a spontaneous fluorogenic conformational change when they hybridize to their targets. Only perfectly complementary targets elicit this response, as hybridization does not occur when the target contains a mismatched nucleotide or a deletion. The probes are particularly suited for monitoring the synthesis of specific nucleic acids in real time. When used in nucleic acid amplification assays, gene detection is homogeneous and sensitive, and can be carried out in a sealed tube. When introduced into living cells, these probes should enable the origin, movement, and fate of specific mRNAs to be traced.
    4. 4
      Qiu, L.; Wu, C.; You, M.; Han, D.; Chen, T.; Zhu, G.; Jiang, J.; Yu, R.; Tan, W. J. Am. Chem. Soc. 2013, 135, 12952– 12955 DOI: 10.1021/ja406252w
      4
      A Targeted, Self-Delivered, and Photocontrolled Molecular Beacon for mRNA Detection in Living Cells
      Qiu, Liping; Wu, Cuichen; You, Mingxu; Han, Da; Chen, Tao; Zhu, Guizhi; Jiang, Jianhui; Yu, Ruqin; Tan, Weihong
      Journal of the American Chemical Society (2013), 135 (35), 12952-12955CODEN: JACSAT; ISSN:0002-7863. (American Chemical Society)
      The spatiotemporal dynamics of specific mRNA mols. are difficult to image and detect inside living cells, and this has been a significant challenge for the chem. and biomedical communities. To solve this problem, we have developed a targeted, self-delivered, and photocontrolled aptamer-based mol. beacon (MB) for intracellular mRNA anal. An internalizing aptamer connected via a double-stranded DNA structure was used as a carrier probe (CP) for cell-specific delivery of the MB designed to signal target mRNA. A light activation strategy was employed by inserting two photolabile groups in the CP sequence, enabling control over the MB's intracellular function. After the probe was guided to the target cell via specific binding of aptamer AS1411 to nucleolin on the cell membrane, light illumination released the MB for mRNA monitoring. Consequently, the MB is able to perform live-cell mRNA imaging with precise spatiotemporal control, while the CP acts as both a tracer for intracellular distribution of the MB before photoinitiation and an internal ref. for mRNA ratiometric detection.
    5. 5
      Seferos, D. S.; Giljohann, D. A.; Hill, H. D.; Prigodich, A. E.; Mirkin, C. A. J. Am. Chem. Soc. 2007, 129, 15477– 15479 DOI: 10.1021/ja0776529
      5
      Nano-Flares: Probes for Transfection and mRNA Detection in Living Cells
      Seferos, Dwight S.; Giljohann, David A.; Hill, Haley D.; Prigodich, Andrew E.; Mirkin, Chad A.
      Journal of the American Chemical Society (2007), 129 (50), 15477-15479CODEN: JACSAT; ISSN:0002-7863. (American Chemical Society)
      We demonstrate that novel oligonucleotide-modified gold nanoparticle probes hybridized to fluorophore-labeled complements can be used as both transfection agents and cellular "nano-flares" for detecting mRNA in living cells. Nano-flares take advantage of the highly efficient fluorescence quenching properties of gold, cellular uptake of oligonucleotide nanoparticle conjugates without the use of transfection agents, and the enzymic stability of such conjugates, thus overcoming many of the challenges to creating sensitive and effective intracellular probes. Nano-flares exhibit high signaling, have low background fluorescence, and are sensitive to changes in the no. of RNA transcripts present in cells.
    6. 6
      Cutler, J. I.; Auyeung, E.; Mirkin, C. A. J. Am. Chem. Soc. 2012, 134, 1376– 1391 DOI: 10.1021/ja209351u
      6
      Spherical Nucleic Acids
      Cutler, Joshua I.; Auyeung, Evelyn; Mirkin, Chad A.
      Journal of the American Chemical Society (2012), 134 (3), 1376-1391CODEN: JACSAT; ISSN:0002-7863. (American Chemical Society)
      A review. A historical perspective of the development of spherical nucleic acid (SNA) conjugates and other three-dimensional nucleic acid nanostructures is provided. This Perspective details the synthetic methods for prepg. them, followed by a discussion of their unique properties and theor. and exptl. models for understanding them. Important examples of technol. advances made possible by their fundamental properties spanning the fields of chem., mol. diagnostics, gene regulation, medicine, and materials science are also presented.
    7. 7
      Visvader, J. E. Nature 2011, 469, 314– 322 DOI: 10.1038/nature09781
      There is no corresponding record for this reference.
    8. 8
      Zhang, H.; Li, F.; Dever, B.; Li, X.; Le, X. C. Chem. Rev. 2013, 113, 2812– 2841 DOI: 10.1021/cr300340p
      8
      DNA-Mediated Homogeneous Binding Assays for Nucleic Acids and Proteins
      Zhang, Hongquan; Li, Feng; Dever, Brittany; Li, Xing-Fang; Le, X. Chris
      Chemical Reviews (Washington, DC, United States) (2013), 113 (4), 2812-2841CODEN: CHREAY; ISSN:0009-2665. (American Chemical Society)
      A review. The present review mainly focuses on recent advances in aptamer beacons, catalytic beacons, in vitro isothermal amplification techniques, DNA nanostructures, and binding-induced DNA assembly for the detection of specific nucleic acid sequences and proteins.
    9. 9
      Zhao, Y.; Chen, F.; Li, Q.; Wang, L.; Fan, C. Chem. Rev. 2015, 115, 12491– 12545 DOI: 10.1021/acs.chemrev.5b00428
      9
      Isothermal Amplification of Nucleic Acids
      Zhao, Yongxi; Chen, Feng; Li, Qian; Wang, Lihua; Fan, Chunhai
      Chemical Reviews (Washington, DC, United States) (2015), 115 (22), 12491-12545CODEN: CHREAY; ISSN:0009-2665. (American Chemical Society)
      Isothermal amplification of nucleic acids is a simple process that rapidly and efficiently accumulates nucleic acid sequences at const. temp. Since the early 1990s, various isothermal amplification techniques have been developed as alternatives to polymerase chain reaction (PCR). These isothermal amplification methods have been used for biosensing targets such as DNA, RNA, cells, proteins, small mols., and ions. The applications of these techniques for in situ or intracellular bioimaging and sequencing have been amply demonstrated. Amplicons produced by isothermal amplification methods have also been utilized to construct versatile nucleic acid nanomaterials for promising applications in biomedicine, bioimaging, and biosensing. The integration of isothermal amplification into microsystems or portable devices improves nucleic acid-based on-site assays and confers high sensitivity. Single-cell and single-mol. analyses have also been implemented based on integrated microfluidic systems. In this review, we provide a comprehensive overview of the isothermal amplification of nucleic acids encompassing work published in the past two decades. First, different isothermal amplification techniques are classified into three types based on reaction kinetics. Then, we summarize the applications of isothermal amplification in bioanal., diagnostics, nanotechnol., materials science, and device integration. Finally, several challenges and perspectives in the field are discussed.
    10. 10
      Dirks, R. M.; Pierce, N. A. Proc. Natl. Acad. Sci. U. S. A. 2004, 101, 15275 DOI: 10.1073/pnas.0407024101
      10
      Triggered amplification by hybridization chain reaction
      Dirks, Robert M.; Pierce, Niles A.
      Proceedings of the National Academy of Sciences of the United States of America (2004), 101 (43), 15275-15278CODEN: PNASA6; ISSN:0027-8424. (National Academy of Sciences)
      We introduce the concept of hybridization chain reaction (HCR), in which stable DNA monomers assemble only upon exposure to a target DNA fragment. In the simplest version of this process, two stable species of DNA hairpins coexist in soln. until the introduction of initiator strands triggers a cascade of hybridization events that yields nicked double helixes analogous to alternating copolymers. The av. mol. wt. of the HCR products varies inversely with initiator concn. Amplification of more diverse recognition events can be achieved by coupling HCR to aptamer triggers. This functionality allows DNA to act as an amplifying transducer for biosensing applications.
    11. 11
      Bi, S.; Yue, S.; Zhang, S. Chem. Soc. Rev. 2017, 46, 4281– 4298 DOI: 10.1039/C7CS00055C
      11
      Hybridization chain reaction: a versatile molecular tool for biosensing, bioimaging, and biomedicine
      Bi, Sai; Yue, Shuzhen; Zhang, Shusheng
      Chemical Society Reviews (2017), 46 (14), 4281-4298CODEN: CSRVBR; ISSN:0306-0012. (Royal Society of Chemistry)
      A review. Developing powerful, simple and low-cost DNA amplification techniques is of great significance to bioanal. and biomedical research. Thus far, many signal amplification strategies have been developed, such as polymerase chain reaction (PCR), rolling circle amplification (RCA), and DNA strand displacement amplification (SDA). In particular, hybridization chain reaction (HCR), a type of toehold-mediated strand displacement (TMSD) reaction, has attracted great interest because of its enzyme-free nature, isothermal conditions, simple protocols, and excellent amplification efficiency. In a typical HCR, an analyte initiates the cross-opening of two DNA hairpins, yielding nicked double helixes that are analogous to alternating copolymers. As an efficient amplification platform, HCR has been utilized for the sensitive detection of a wide variety of analytes, including nucleic acids, proteins, small mols., and cells. In recent years, more complicated sets of monomers have been designed to develop nonlinear HCR, such as branched HCR and even dendritic systems, achieving quadratic and exponential growth mechanisms. In addn., HCR has attracted enormous attention in the fields of bioimaging and biomedicine, including applications in fluorescence in situ hybridization (FISH) imaging, live cell imaging, and targeted drug delivery. In this review, we introduce the fundamentals of HCR and examine the visualization and anal. techniques for HCR products in detail.
    12. 12
      Chen, Y.; Xu, J.; Su, J.; Xiang, Y.; Yuan, R.; Chai, Y. Anal. Chem. 2012, 84, 7750– 7755 DOI: 10.1021/ac3012285
      12
      In situ hybridization chain reaction amplification for universal and highly sensitive electrochemiluminescent detection of DNA
      Chen, Ying; Xu, Jin; Su, Jiao; Xiang, Yun; Yuan, Ruo; Chai, Yaqin
      Analytical Chemistry (Washington, DC, United States) (2012), 84 (18), 7750-7755CODEN: ANCHAM; ISSN:0003-2700. (American Chemical Society)
      In this work, we describe a new universal and highly sensitive strategy for electrochemiluminescent (ECL) detection of sequence specific DNA at the femtomolar level via in situ hybridization chain reaction (HCR) signal amplification. The DNA capture probes are self-assembled on a gold electrode. The presence of the target DNA and two hairpin helper DNAs leads to the formation of extended dsDNA polymers through HCR on the electrode surface. The in situ, HCR-generated dsDNA polymers cause the intercalation of numerous ECL indicators (Ru(phen)32+) into the dsDNA grooves, resulting in significantly amplified ECL signal output. The proposed strategy combines the amplification power of the DNA HCR and the inherent high sensitivity of the ECL technique and enables low femtomolar detection of sequence specific DNA. The developed strategy also shows high selectivity against single-base mismatch sequences, which makes our new universal and highly sensitive HCR-based method a useful addn. to the amplified DNA detection arena.
    13. 13
      Cheglakov, Z.; Cronin, T. M.; He, C.; Weizmann, Y. J. Am. Chem. Soc. 2015, 137, 6116– 6119 DOI: 10.1021/jacs.5b01451
      13
      Live cell microRNA imaging using cascade hybridization reaction
      Cheglakov, Zoya; Cronin, Timothy M.; He, Chuan; Weizmann, Yossi
      Journal of the American Chemical Society (2015), 137 (19), 6116-6119CODEN: JACSAT; ISSN:0002-7863. (American Chemical Society)
      Recent advances in RNA research have posed new directives in biol. and chem. to uncover the complex roles of ribonucleic acids in cellular processes. Innovative techniques to visualize native RNAs, particularly, short, low-abundance RNAs in live cells, can dramatically impact current research on the roles of RNAs in biol. Herein, we report a novel method for real-time, microRNA imaging inside live cells based on programmable oligonucleotide probes, which self-assemble through the Cascade Hybridization Reaction (CHR).
    14. 14
      Jiang, Y.; Li, B.; Milligan, J. N.; Bhadra, S.; Ellington, A. D. J. Am. Chem. Soc. 2013, 135, 7430 DOI: 10.1021/ja4023978
      14
      Real-time detection of isothermal amplification reactions with thermostable catalytic hairpin assembly
      Jiang, Yu; Li, Bingling; Milligan, John N.; Bhadra, Sanchita; Ellington, Andrew D.
      Journal of the American Chemical Society (2013), 135 (20), 7430-7433CODEN: JACSAT; ISSN:0002-7863. (American Chemical Society)
      Catalytic hairpin assembly (CHA) is an enzyme-free amplification method that has previously proven useful in amplifying and transducing signals at the terminus of nucleic acid amplification reactions. Here, for the first time, we engineered CHA to be thermostable from 37 to 60° and in consequence have generalized its application to the real-time detection of isothermal amplification reactions. CHA circuits were designed and optimized for both high- and low-temp. rolling circle amplification (RCA) and strand displacement amplification (SDA). The resulting circuits not only increased the specificity of detection but also improved the sensitivity by as much as 25- to 10000-fold over comparable real-time detection methods. These methods have been condensed into a set of general rules for the design of thermostable CHA circuits with high signals and low noise.
    15. 15
      Zhang, D. Y.; Turberfield, A. J.; Yurke, B.; Winfree, E. Science 2007, 318, 1121– 1125 DOI: 10.1126/science.1148532
      15
      Engineering Entropy-Driven Reactions and Networks Catalyzed by DNA
      Zhang, David Yu; Turberfield, Andrew J.; Yurke, Bernard; Winfree, Erik
      Science (Washington, DC, United States) (2007), 318 (5853), 1121-1125CODEN: SCIEAS; ISSN:0036-8075. (American Association for the Advancement of Science)
      Artificial biochem. circuits are likely to play as large a role in biol. engineering as elec. circuits have played in the engineering of electromech. devices. Toward that end, nucleic acids provide a designable substrate for the regulation of biochem. reactions. However, it has been difficult to incorporate signal amplification components. We introduce a design strategy that allows a specified input oligonucleotide to catalyze the release of a specified output oligonucleotide, which in turn can serve as a catalyst for other reactions. This reaction, which is driven forward by the configurational entropy of the released mol., provides an amplifying circuit element that is simple, fast, modular, composable, and robust. We have constructed and characterized several circuits that amplify nucleic acid signals, including a feedforward cascade with quadratic kinetics and a pos. feedback circuit with exponential growth kinetics.
    16. 16
      Wu, C.; Cansiz, S.; Zhang, L.; Teng, I.; Qiu, L.; Li, J.; Liu, Y.; Zhou, C.; Hu, R.; Zhang, T.; Cui, C.; Cui, L.; Tan, W. J. Am. Chem. Soc. 2015, 137, 4900– 4903 DOI: 10.1021/jacs.5b00542
      16
      A nonenzymatic hairpin DNA cascade reaction provides high signal gain of mRNA imaging inside live cells
      Wu, Cuichen; Cansiz, Sena; Zhang, Liqin; Teng, I-Ting; Qiu, Liping; Li, Juan; Liu, Yuan; Zhou, Cuisong; Hu, Rong; Zhang, Tao; Cui, Cheng; Cui, Liang; Tan, Weihong
      Journal of the American Chemical Society (2015), 137 (15), 4900-4903CODEN: JACSAT; ISSN:0002-7863. (American Chemical Society)
      Enzyme-free signal amplification has enabled sensitive in vitro detection of biomols. such as proteins and nucleic acids. However, monitoring targets of interest in live cells via enzyme-free amplification is still challenging, esp. for analytes with low concns. To the best of our knowledge, this paper reports the first attempt to perform mRNA imaging inside live cells, using a nonenzymic hairpin DNA cascade reaction for high signal gain, termed a hairpin DNA cascade amplifier (HDCA). In conventional nucleic acid probes, such as linear hybridization probes, mRNA target signaling occurs in an equiv. reaction ratio (1:1), whereas, in HDCA, one mRNA target is able to yield multiple signal outputs (1:m), thus achieving the goal of signal amplification for low-expression mRNA targets. Moreover, the recycled mRNA target in the HDCA serves as a catalyst for the assembly of multiple DNA duplexes, generating the fluorescent signal of reduced MnSOD mRNA expression, thus indicating amplified intracellular imaging. This programmable cascade reaction presents a simple and modular amplification mechanism for intracellular biomarkers of interest, providing a significant boost to the search for clues leading to the accurate identification and effective treatment of cancers.
    17. 17
      Li, L.; Feng, J.; Liu, H.; Li, Q.; Tong, L.; Tang, B. Chem. Sci. 2016, 7, 1940– 1945 DOI: 10.1039/C5SC03909F
      17
      Two-color imaging of microRNA with enzyme-free signal amplification via hybridization chain reactions in living cells
      Li, Lu; Feng, Jie; Liu, Haiyun; Li, Qingling; Tong, Lili; Tang, Bo
      Chemical Science (2016), 7 (3), 1940-1945CODEN: CSHCCN; ISSN:2041-6520. (Royal Society of Chemistry)
      In situ imaging of miRNA in living cells could facilitate the monitoring of the dynamic expression and distribution of miRNA and research on miRNA-related cellular processes and diseases. Given the low expression levels and even down-regulation of cellular miRNA that is assocd. with some diseases, amplification strategies are imperative for intracellular miRNA imaging. The present paper proposes a non-destructive amplification strategy for use in living cells. This amplification strategy utilizes the enzyme-free hybridization chain reaction (HCR) with graphene oxide (GO) as a carrier to image cellular miRNA. The resulting signal amplification provides excellent recognition and signal enhancement of specific miRNAs in living cells. As the fluorescence quencher and probe carrier, GO enables activation of the signal switch and effective intracellular delivery of amplification reagents. This new imaging method realizes simple, sensitive and non-destructive signal amplification of miRNA in living cells and has an ability to simultaneously image two types of miRNA in the same cell. This method supplies accurate information regarding cellular miRNA-related biol. events and provides a new tool for highly sensitive and simultaneous imaging of multiple low-level biomarkers, thereby improving the accuracy of early disease diagnosis.
    18. 18
      Wu, Z.; Liu, G. Q.; Yang, X. L.; Jiang, J. J. Am. Chem. Soc. 2015, 137, 6829 DOI: 10.1021/jacs.5b01778
      18
      Electrostatic nucleic acid nanoassembly enables hybridization chain reaction in living cells for ultrasensitive mRNA imaging
      Wu, Zhan; Liu, Gao-Qin; Yang, Xiao-Li; Jiang, Jian-Hui
      Journal of the American Chemical Society (2015), 137 (21), 6829-6836CODEN: JACSAT; ISSN:0002-7863. (American Chemical Society)
      Efficient approaches for intracellular delivery of nucleic acid reagents to achieve sensitive detection and regulation of gene and protein expressions are essential for chem. and biol. We develop a novel electrostatic DNA nanoassembly that, for the first time, realizes hybridization chain reaction (HCR), a target-initiated alternating hybridization reaction between two hairpin probes, for signal amplification in living cells. The DNA nanoassembly has a designed structure with a core gold nanoparticle, a cationic peptide interlayer, and an electrostatically assembled outer layer of fluorophore-labeled hairpin DNA probes. It is shown to have high efficiency for cellular delivery of DNA probes via a unique endocytosis-independent mechanism that confers a significant advantage of overcoming endosomal entrapment. Moreover, electrostatic assembly of DNA probes enables target-initialized release of the probes from the nanoassembly via HCR. This intracellular HCR offers efficient signal amplification and enables ultrasensitive fluorescence activation imaging of mRNA expression with a picomolar detection limit. The results imply that the developed nanoassembly may provide an invaluable platform in low-abundance biomarker discovery and regulation for cell biol. and theranostics.
    19. 19
      Tang, Y.; Zhang, X.; Tang, L.; Yu, R.; Jiang, J. Anal. Chem. 2017, 89, 3445– 3451 DOI: 10.1021/acs.analchem.6b04312
      19
      In Situ Imaging of Individual mRNA Mutation in Single Cells Using Ligation-Mediated Branched Hybridization Chain Reaction (Ligation-bHCR)
      Tang, Ying; Zhang, Xiao-Li; Tang, Li-Juan; Yu, Ru-Qin; Jiang, Jian-Hui
      Analytical Chemistry (Washington, DC, United States) (2017), 89 (6), 3445-3451CODEN: ANCHAM; ISSN:0003-2700. (American Chemical Society)
      Ultrasensitive and specific in situ imaging of gene expression is essential for mol. medicine and clin. theranostics. The authors develop a novel fluorescence in situ hybridization (FISH) strategy based on a new branched hybridization chain reaction (bHCR) for efficient signal amplification in FISH assay and a ligase-mediated discrimination for specific mutation detection. To the knowledge, this is the first time that HCR has been realized for mutation detection in FISH assay. In vitro assay shows that the ligation-bHCR strategy affords high specificity in discriminating single-nucleotide variation in mRNA, and generates a highly branched polymeric product that confers more efficient amplification or better sensitivity than HCR. Imaging anal. reveals that ligation-bHCR generates highly bright spot-like signals for localization of individual mRNA mols., and spot signals of different colors are highly specific in genotyping point mutation of individual mRNA. Moreover, this strategy is shown to have the potential for quant. i.m.-aging of the expression of mRNA at the single cell level. Therefore, this strategy may provide a new promising paradigm in developing highly sensitive and specific FISH methods for various diagnostic and research applications.
    20. 20
      Xuan, F.; Hsing, I. M. J. Am. Chem. Soc. 2014, 136, 9810– 9813 DOI: 10.1021/ja502904s
      20
      Triggering hairpin-free chain-branching growth of fluorescent DNA dendrimers for nonlinear hybridization chain reaction
      Xuan, Feng; Hsing, I.-Ming
      Journal of the American Chemical Society (2014), 136 (28), 9810-9813CODEN: JACSAT; ISSN:0002-7863. (American Chemical Society)
      The authors present a nonlinear hybridization chain reaction (HCR) system in which a trigger DNA initiates self-sustained assembly of quenched double-stranded substrates into fluorescent dendritic nanostructures. During the process, an increasing no. of originally sequestered trigger sequences labeled with fluorescent reporters are freed up from quenched substrates, leading to chain-branching growth of the assembled DNA dendrimers and an exponential increase in the fluorescence intensity. The triggered assembly behavior was examd. by PAGE anal., and the morphologies of the grown dendrimers were verified by AFM imaging. The exponential kinetics of the fluorescence accumulation was also confirmed by time-dependent fluorescence spectroscopy. This method adopts a simple sequence design strategy, the concept of which could be adapted to program assembly systems with higher-order growth kinetics.
    21. 21
      Bi, S.; Chen, M.; Jia, X.; Dong, Y.; Wang, Z. Angew. Chem., Int. Ed. 2015, 54, 8144– 8148 DOI: 10.1002/anie.201501457
      There is no corresponding record for this reference.
    22. 22
      Chandran, H.; Rangnekar, A.; Shetty, G.; Schultes, E. A.; Reif, J. H.; LaBean, T. H. Biotechnol. J. 2013, 8, 221– 227 DOI: 10.1002/biot.201100499
      22
      An autonomously self-assembling dendritic DNA nanostructure for target DNA detection
      Chandran, Harish; Rangnekar, Abhijit; Shetty, Geetha; Schultes, Erik A.; Reif, John H.; LaBean, Thomas H.
      Biotechnology Journal (2013), 8 (2), 221-227CODEN: BJIOAM; ISSN:1860-6768. (Wiley-VCH Verlag GmbH & Co. KGaA)
      There is a growing need for sensitive and reliable nucleic acid detection methods that are convenient and inexpensive. Responsive and programmable DNA nanostructures have shown great promise as chem. detection systems. Here, we describe a DNA detection system employing the triggered self-assembly of a novel DNA dendritic nanostructure. The detection protocol is executed autonomously without external intervention. Detection begins when a specific, single-stranded target DNA strand (T) triggers a hybridization chain reaction (HCR) between two, distinct DNA hairpins (α and β). Each hairpin opens and hybridizes up to two copies of the other. In the absence of T, α and β are stable and remain in their poised, closed-hairpin form. In the presence of T, α hairpins are opened by toe-hold mediated strand-displacement, each of which then opens and hybridizes two β hairpins. Likewise, each opened β hairpin can open and hybridize two α hairpins. Hence, each layer of the growing dendritic nanostructure can in principle accommodate an exponentially increasing no. of cognate mols., generating a high mol. wt. nanostructure. This HCR system has minimal sequence constraints, allowing reconfiguration for the detection of arbitrary target sequences. Here, we demonstrate detection of unique sequence identifiers of HIV and Chlamydia pathogens.
    23. 23
      Wei, J.; Gong, X.; Wang, Q.; Pan, M.; Liu, X.; Liu, J.; Xia, F.; Wang, F. Chem. Sci. 2018, 9, 52 DOI: 10.1039/C7SC03939E
      23
      Construction of an autonomously concatenated hybridization chain reaction for signal amplification and intracellular imaging
      Wei, Jie; Gong, Xue; Wang, Qing; Pan, Min; Liu, Xiaoqing; Liu, Jing; Xia, Fan; Wang, Fuan
      Chemical Science (2018), 9 (1), 52-61CODEN: CSHCCN; ISSN:2041-6520. (Royal Society of Chemistry)
      Biomol. self-assembly has spurred substantial research efforts for the development of low-cost point-of-care diagnostics. Herein, we introduce an isothermal enzyme-free concatenated hybridization chain reaction (C-HCR), in which the output of the upstream hybridization chain reaction (HCR-1) layer acts as an intermediate input to activate the downstream hybridization chain reaction (HCR-2) layer. The initiator motivates HCR-1 through the autonomous cross-opening of two functional DNA hairpins, yielding polymeric dsDNA nanowires composed of numerous tandem triggers T as output of the primary sensing event. The reconstituted amplicon T then initiates HCR-2 and transduces the analyte recognition into an amplified readout, originating from the synergistic effect between HCR-1 and HCR-2 layers. Native gel electrophoresis, atom force microscopy (AFM) and fluorescence spectra revealed that C-HCR mediated the formation of frond-like branched dsDNA nanowires and the generation of an amplified FRET signal. As a versatile and robust amplification strategy, the unpreceded C-HCR can discriminate DNA analyte from its mutants with high accuracy and specificity. By incorporating an auxiliary sensing module, the integrated C-HCR amplifier was further adapted for highly sensitive and selective detection of microRNA (miRNA), as a result of the hierarchical and sequential hybridization chain reactions, in human serum and even living cells through an easy-to-integrate "plug-and-play" procedure. In addn., the C-HCR amplifier was successfully implemented for intracellular miRNA imaging by acquiring an accurate and precise signal localization inside living cells, which was esp. suitable for the ex situ and in situ amplified detection of trace amts. of analyte. The C-HCR amplification provides a comprehensive and smart toolbox for highly sensitive detection of various biomarkers and thus should hold great promise in clin. diagnosis and assessment. The infinite layer of multilayered C-HCR is anticipated to further strengthen the amplification capacity and reliability (anti-invasion performance) of intracellular imaging approach, which is of great significance for its bioanal. applications.
    24. 24
      Chi, Y.; Wang, X.; Yang, Y.; Zhang, C.; Ertl, H. C.; Zhou, D. Mol. Ther.--Nucleic Acids 2014, 3, e208 DOI: 10.1038/mtna.2014.59
      24
      Survivin-targeting Artificial MicroRNAs Mediated by Adenovirus Suppress Tumor Activity in Cancer Cells and Xenograft Models
      Chi, Yudan; Wang, Xiang; Yang, Yong; Zhang, Chao; Ertl, Hildegund C. J.; Zhou, Dongming
      Molecular Therapy--Nucleic Acids (2014), 3 (11), e208CODEN: MTAOC5; ISSN:2162-2531. (Nature Publishing Group)
      Survivin is highly expressed in most human tumors and fetal tissue, and absent in terminally differentiated cells. It promotes tumor cell proliferation by neg. regulating cell apoptosis and facilitating cell division. Survivin's selective expression pattern suggests that it might be a suitable target for cancer therapy, which would promote death of transformed but not normal cells. This was tested using artificial microRNAs (amiRNAs) targeting survivin. After screening, two effective amiRNAs, which knocked down survivin expression, were identified and cloned into a replication-defective adenoviral vector. Tumor cells infected with the recombinant vector downregulated expression of survivin and underwent apoptotic cell death. Further studies showed that apoptosis was assocd. with increases in caspase 3 and cleaved Poly (ADP-ribose) polymerase, and activation of the p53 signaling pathway. Furthermore, amiRNA treatment caused blockade of mitosis and cell cycle arrest at the G2/M phase. In vivo, survivin-targeting amiRNAs expressed by adenoviral vectors effectively delayed growth of hepatocellular and cervical carcinomas in mouse xenograft models. These results indicate that silencing of survivin by amiRNA has potential for treatment of cancer.
    25. 25
      Peng, X. H.; Cao, Z. H.; Xia, J. T.; Carlson, G. W.; Lewis, M. M.; Wood, W. C.; Yang, L. Cancer Res. 2005, 65, 1909– 1917 DOI: 10.1158/0008-5472.CAN-04-3196
      There is no corresponding record for this reference.
    26. 26
      Yamanaka, K.; Nakahara, T.; Yamauchi, T.; Kita, A.; Takeuchi, M.; Kiyonaga, F.; Kaneko, N.; Sasamata, M. Clin. Cancer Res. 2011, 17, 5423– 31 DOI: 10.1158/1078-0432.CCR-10-3410
      26
      Antitumor Activity of YM155, a Selective Small-Molecule Survivin Suppressant, Alone and in Combination with Docetaxel in Human Malignant Melanoma Models
      Yamanaka, Kentaro; Nakahara, Takahito; Yamauchi, Tomohiro; Kita, Aya; Takeuchi, Masahiro; Kiyonaga, Fumiko; Kaneko, Naoki; Sasamata, Masao
      Clinical Cancer Research (2011), 17 (16), 5423-5431CODEN: CCREF4; ISSN:1078-0432. (American Association for Cancer Research)
      Purpose: Aggressive cell growth and chemoresistance are notorious obstacles in melanoma therapy. Accumulating evidence suggests that survivin is preferentially expressed in cancer cells and plays a crucial role in cell division and apoptosis dysfunction. Here, we evaluated the therapeutic potential of YM155, a selective survivin suppressant, alone and in combination with docetaxel using human melanoma models. Exptl. Design: A375 and SK-MEL-5 human malignant melanoma cells were treated with siRNA, YM155, and/or docetaxel, and cell viability, mRNA and protein expression levels, cell-cycle distribution, and immunohistochem. staining were then evaluated. Furthermore, the efficacy of YM155 combined with docetaxel was further examd. in established xenograft models. Results: Survivin suppression was sufficient to induce spontaneous apoptosis of melanoma cells. YM155 showed nanomolar antiproliferative effects and induced tumor regression in established melanoma xenograft models. Docetaxel showed antitumor activity against melanoma cells, although it also induced survivin upregulation and G2/M mitotic arrest; however, cotreatment with YM155 decreased survivin expression below basal levels. Combination treatment of YM155 and docetaxel induced a greater rate of apoptosis than the sum of the single-treatment rates and promoted tumor regression without enhanced body wt. loss in the melanoma xenograft models. Conclusions: Survivin is responsible for the inherent low levels of spontaneous apoptosis in melanoma cells. The concomitant combination of YM155 with docetaxel diminished the accumulation of survivin in G2/M mitotic arrest, and induced more intense apoptosis compared with each single treatment. YM155 in combination with docetaxel is well tolerated and shows greater efficacy than either agent alone in mouse xenograft models. Clin Cancer Res; 17(16); 5423-31.

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