ACS Publications. Most Trusted. Most Cited. Most Read
Quick View

OR SEARCH CITATIONS

My Activity
Publications
CONTENT TYPES

Figure 1Loading Img
Download Hi-Res ImageDownload to MS-PowerPointCite This:J. Am. Chem. Soc. 2017, 139, 35, 12117-12120
RETURN TO ISSUEPREVCommunicationNEXT

Magnetic Field-Activated Sensing of mRNA in Living Cells

View Author Information
† Department of Chemistry and Biomolecular Science, Clarkson University, Potsdam, New York 13699-5810, United States
§ Nanostructured Materials Lab, University of Georgia, Athens, Georgia 30602, United States
‡ Department of Biology, Clarkson University, Potsdam, New York 13699-5810, United States
∥ Department of Microbiology, Immunology, and Molecular Genetics, University of Texas Health Science Center at San Antonio, 7703 Floyd Curl Drive, San Antonio, Texas 78229-3900, United States
# Chemistry Department, University of Central Florida, 4000 Central Florida Boulevard, Orlando, Florida 32816-2366, United States
⊥ Laboratory of Solution Chemistry of Advanced Materials and Technologies, ITMO University, Lomonosova St. 9, 191002 St. Petersburg, Russian Federation
*[email protected]
*[email protected]
*[email protected]
Cite this: J. Am. Chem. Soc. 2017, 139, 35, 12117–12120
Publication Date (Web):August 17, 2017
https://doi.org/10.1021/jacs.7b06022

Copyright © 2017 American Chemical Society. This publication is licensed under these Terms of Use.

Request reuse permissions
  • Open Access

Article Views

9333

Altmetric

-

Citations

43
LEARN ABOUT THESE METRICS
PDF (2 MB)
Get e-Alerts
Supporting Info (2)»
SUBJECTS:

Abstract

High Resolution Image
Download MS PowerPoint Slide

Detection of specific mRNA in living cells has attracted significant attention in the past decade. Probes that can be easily delivered into cells and activated at the desired time can contribute to understanding translation, trafficking and degradation of mRNA. Here we report a new strategy termed magnetic field-activated binary deoxyribozyme (MaBiDZ) sensor that enables both efficient delivery and temporal control of mRNA sensing by magnetic field. MaBiDZ uses two species of magnetic beads conjugated with different components of a multicomponent deoxyribozyme (DZ) sensor. The DZ sensor is activated only in the presence of a specific target mRNA and when a magnetic field is applied. Here we demonstrate that MaBiDZ sensor can be internalized in live MCF-7 breast cancer cells and activated by a magnetic field to fluorescently report the presence of specific mRNA, which are cancer biomarkers.

The development of green fluorescent protein (GFP) for intracellular imaging of specific proteins was acknowledged by a Nobel Prize in Chemistry in 2008. (1) Imaging of specific mRNA inside individual cells is another important task that can contribute to both understanding of mRNA processing and to probing the functions of recently discovered noncoding RNAs. (2) A great variety of approaches for targeted sensing of mRNA in live cells has been proposed in recent years including aptamer-protein systems (e.g., MS2 system), (3) aptamer-dye systems (e.g., spinach aptamer), (4) nucleic acid templated chemical reactions, (5) adjacent hybridization probes, (6) molecular beacon (MB) probes (7) and nanoparticle-based approaches, (8, 9) among others. (10) However, the delivery of the probes or expression of fluorescent species within genetically modified cells requires hours of incubation. Development of a probe that could be activated with a remotely applied physical stimulus would enable activation of sensing and quantification of mRNA in cells at the desired time point. Caged MB probes have been suggested for light-activated detection, which can potentially enable temporal control of sensing. (11) However, caged MB probes produce high background fluorescence (11b, 12) and require invasive delivery of the probe inside cells. (13) Moreover, light-dependent activation may result in either incomplete probe activation or photodamage to living cells. Therefore, no efficient approaches for instant, remotely activated sensing of mRNA inside cells are available to date.

Nanomagnetic actuation (14) (activation of biomolecular species bound to magnetic nanoparticles in the presence of an externally applied magnetic field) elegantly addresses the common issues faced by other comparative techniques for the remote sensing and actuation of intracellular processes. Indeed, owing to its high precision and accuracy, the coupling of a magnetic field to a biomolecule-conjugated magnetic nanoparticle has been applied to several areas of biomedical science: for the investigation of cell mechanical properties, (15) mechanosensitive ion channel signaling pathways, (16) and for targeted activation of specific ion channels. (17)

Here we report a new RNA sensing technology based on the principles of nanomagnetic actuation, magnetic field-activated binary deoxyribozyme (MaBiDZ), which enables sensing of a specific mRNA analyte via application of a magnetic field in a remote and noninvasive manner. The technology takes advantage of magnetic beads (MaB) coupled to a binary deoxyribozyme (BiDZ) probe (Scheme 1A), developed earlier. (18) BiDZ consists of three components: the analyte binding arms (DZa and DZb) and a fluorogenic reporter substrate (F-sub). F-sub is an oligonucleotide strand composed of a fluorophore and quencher conjugated to the opposite sides of the cleavage site. DZa and DZb can hybridize to a specific DNA or RNA analyte and form the DZ catalytic core, which cleaves F-sub, thus resulting in separation of the fluorophore and quencher followed by fluorescent signaling. Important advantages of BiDZ over other hybridization probes is its improved sensitivity, single mismatch selectivity at ambient temperatures, simple design, and low cost. (18) In this work, we took advantage of the modular design and high sensitivity of the BiDZ probe for the development of MaBiDZ, a novel magnetic field-activated switch for real time mRNA sensing in live cells.

Scheme 1

Scheme 1. Principle of Magnetic Field-Activated Deoxyribozyme Sensora
High Resolution Image
Download MS PowerPoint Slide

Scheme a(A) Binary deoxyribozyme sensor (BiDZ) as reported earlier. (18) DNA strands DZa and DZb hybridize to adjacent position of analyte and form deoxyribozyme catalytic core, which cleaves fluorogenic F-sub and increases sample fluorescence. (B) MaBiDZ developed in this study. Magnetic bead (MaB1)-bound DZa forms a catalytic core with DZb in the presence of analyte. The activated sensor produces signal only when: (i) second species of magnetic beads, MaB2 carrying F-sub is present and (ii) magnetic field that aggregates MaB1 and MaB2 is applied. See the DNA sequences in Table 1.

The magnetic switch consists of two species of 100 nm magnetic beads (MaB), MaB1 and MaB2 (Scheme 1B). MaB is composed of a 15 nm iron oxide (Fe3O4) superparamagnetic core encased in a silica shell. The shell is modified with a grafted polymeric brush of a block copolymer PAA-b-PEGMA composed of poly(acrylic acid) (PAA) and a polymer of poly(ethylene glycol) methyl ether acrylate (PEGMA). The MaB cores have a saturation magnetization value (47 emu/g), (19) which is sufficient for effective utilization of magnetic force. The DNA strands are conjugated to MaB via the polymeric brush using a flexible linker, which is known to improve biocompatibility, facilitate intracellular delivery and prevent nanoparticle aggregation in the absence of a magnetic field, (20) whereas the flexible linker allows mobility of the BiDZ arms.

MaBiDZ consists of the DZb strand, MaB1 conjugated with DZa and MaB2 conjugated with DNA hook strand complementary to F-sub (Scheme 1B, see SI for details of the conjugation procedure). F-sub is incubated with the Hook-MaB2 conjugate, which is then rinsed to remove unbound F-sub. A DNA or RNA analyte hybridized to DZa and DZb strands enables formation of the DZ catalytic core. The catalytic core does not produce the fluorescent signal unless hybridized with F-sub. Application of an external magnetic field induces aggregation of the MaB1 and MaB2, thus bringing the activated BiDZ sensor in close proximity to F-sub, which is followed by F-sub cleavage and amplification of fluorescent signal. Though the 3D motion of MaBiDZ may be restricted under a magnetic field, both the flexible linker and large particle size allow a greater degree of contact points between the two DZ species. To the best of our knowledge this is the first strategy that allows activation of a hybridization sensor by a magnetic field. Another important advantage of this approach is the low background fluorescence due to the low concentration of the F-sub in solution, in comparison with the BiDZ detection (Scheme 1A). Indeed, the amount of F-sub attached to the beads is much lower than that used by BiDZ sensor (typically 200 nM). However, when MaB1 and MaB2 are aggregated, the local concentration of F-sub near the activated sensor is high.

For the proof-of-concept study, we chose to target Twist mRNA. Twist is a helix–loop–helix transcription factor whose overexpression has been shown to contribute to metastasis by promoting an epithelial-mesenchymal transition. (21) Thus, an intracellular sensor that can fluorescently report Twist mRNA levels would be useful to assess metastatic potential of cells in clinical applications. We first optimized the performance of the sensor in in vitro experiments using a synthetic DNA analyte with the sequence of Twist mRNA (see Twist sequence in Table 1).

Table 1. Oligonucleotides Used in the Study
NameaSequences
F-sub5′-CGGT ACA TTG TAG AAG TT AAG GTTFAM TCC TCg uCC CTG GGC A-BHQ1
Twist5′-TAGT GGG ACG CGG ACA TGG ACC AGG CCC CCT CCA TCC TCC AGA CCG AGA AGG CGT AGC TGA GCC GCT CGT GAG CCA CAT AGC TGC A
DZa5′-NH2/AAA AAA AAA AAA AAA AAA AAC GAG CGG CTC AGC TAC GCC T AC AAC CGA GAG AGG AAA C
DZb5′-CCA GGG A GG CTA GCT TCT CGG TCT GGA GGA TGG AG
Hook5′-NH2/AAA AAA AAA AAA AAA AAA AA/iSp9/AAC TTC TAC AAT GTA CCG
a

iSp9 - triethylene glycol linker; FAM attached to the DNA is a fluorescein derivative; BHQ1 - “Black Hole Quencher” is a fluorescence quencher; ribonucleotides are in low case.

The results of in vitro studies demonstrated a near 2-fold enhancement of fluorescent signal when MaBiDZ is switched ON in the presence of the magnetic field compared to the OFF state, for which the signal does not change over time (Figure 1A). Importantly, the signal remained at the background level in the absence of an applied magnetic field (Figure 1A, (c)) and in the absence of analyte (Figure 1A, (b)). Furthermore, the signal response to a noncomplementary target is similar to that of the background fluorescence (see SI for Figure SI5). The results also demonstrate faster activation of MaBiDZ compared to BiDZ, (Figure 1A, compare slopes of lines e and d). Time dependent profiles demonstrate that unlike BiDZ, MaBiDZ does not demonstrate time dependence of fluorescent response, but produces maximum signal within 30 min. These data demonstrate important advantages of MaBiDZ system in comparison with BiDZ: (1) it responds faster upon activation by magnetic field and reaches maximum signal in shorter time, due to, presumably, higher local concentration of F-sub. MaBiDZ’s response does not increase over longer incubation time due to the shortage of F-sub supply, which is limited by contact area between MaB1 and MaB2. Thus, an important feature of the MaBiDZ system is activation at the desired time. (2) The two-probe system allows measurement of Twist in a controlled fashion by separating F-sub and DZa, lowering the background signal generated.

Figure 1

Figure 1. Comparison of in vitro fluorescent response of BiDZ and MaBiDZ sensor systems. (A) Time dependent response of BiDZ and MaBiDZ sensor: (a) without F-sub, (b) without synthetic Twist (see Table1) analyte, (c) response of MaBiDz without magnet applied, (d) response of BiDZ, and (e) MaBiDZ activated with magnetic field in the presence of 1 nM synthetic Twist analyte (see SI for concentrations of all other components of the BiDZ and MaBiDZ probe.) (B) Response of MaBiDZ (d, e, f) compared to BiDZ (a, b, c) in the presence of different concentrations of Twist analyte after 30 (a,d), 60 (b,e) or 120 (c,f) min. All error bars are the result of three independent measurements; some bars are not visible because they are smaller than the labels for the experimental points.

High Resolution Image
Download MS PowerPoint Slide

Next we chose to test the sensing ability of MaBiDZ ex vivo in mammalian cell culture. We chose the MCF-7 breast cancer cell line, and human cervical epithelial cells (HCX) isolated from human tissue, which express high (22) and lower levels (23) of Twist mRNA, respectively. Our first aim was to compare the fluorescent response of MaBiDZ in MCF-7 cells with and without exposure to a magnetic field (ON and OFF states, respectively) using confocal laser scanning microscopy (CLSM). Cells were incubated with MaBiDZ at a 40 μg/mL concentration (within the none-toxicity concentration range, see Figure SI9) for 4 h and monitored with CLSM every 30 min. Cells exposed to a magnetic field demonstrated a highly fluorescent response compared to those without a magnetic field (compare green fluorescence in panels A and C, Figure 2). Next, we tested MaBiDZ in its ability to detect different levels of mRNA. CLSM images demonstrated higher fluorescent responses in MCF-7 (panel A and C) than in HCX (panel B and D) cells consistent with the reported differences in Twist mRNA levels. (22, 23)

Figure 2

Figure 2. Intracellular testing of MaBiDZ sensor. CLSM images of (A) Twist-overexpressing MCF-7 cancer cells treated with MaBiDZ sensor with magnetic field applied and (C) no magnetic field applied. Analogously treated cervical epithelial cells (expressing low levels of Twist) with (B) magnetic field applied and (D) without magnetic field. Images were taken after 2.5 h of incubation time. Nuclei are stained with Hoechst nuclear stain and visualized with 408 nm laser. Surfaces are stained with anti-epithelial cell adhesion molecule (EpCAM) antibody and visualized with a 635 nm laser. Fluorescence from the MaBiDZ probe is visualized with the 488 nm laser. Corresponding flow cytometry data are shown as insets below each image. The gates on flow cytometry plots indicate percent of EpCAM positive cells with low and high MaBiDZ fluorescence. The number of internalized particles was estimated to be ca. 1 × 106 MaBiDZ per cell (see Figure SI11). Scale bar is 20 μm.

High Resolution Image
Download MS PowerPoint Slide

To quantify the intracellular signaling of the MaBiDZ probe, we examined large population of cells treated with probes using flow cytometry. This method eliminates variations that can be observed using CLSM, which only permits the examination of a small fraction of cells. Flow cytometry results (Figure 2, insets) show that MCF-7 cells treated with MaBiDZ and a magnetic field (ON state) exhibited 4 times greater fluorescence than MaBiDZ-treated MCF-7 cells without a magnetic field (OFF state), thus confirming the magnetic field-dependent switch-like effect of this system (compare insets in Figure 2A,C). When compared to the control noncancerous HCX cells, MCF-7 cells exhibited a 20-fold fluorescence enhancement (compare panel A with B, in Figure 2). It is important to note that significant signaling was apparent after only 2.5 h, as opposed to a previous technique that required an incubation of 12 h before a signal could be detected. (9) To demonstrate the low background of MaBiDZ, we incubated MaB2 (bound to F-sub) alone in MCF-7 cells (see SI, Figure SI6). A signal enhancement was not observed, confirming that MaB-attachment protects F-sub from nuclease-induced cleavage, which would result in high background fluorescence. Earlier, a similar effect was observed for gold nanoparticle-attached fluorescent probes. (8) The fluorescence data from CLSM and flow cytometry measurements of whole cells was validated using fluorescence data of cell lysates (see SI, Figure SI10). This data was in good agreement with measurements of Twist levels from whole cells.

Our next aim was to investigate the mechanisms that promote the observed signaling efficiency and enhancement of MaBiDz within the cell. We hypothesized that the magnetic field plays a role in enhancing cellular entry and intracellular transport kinetics, based on previous reports. (24) To investigate this, we examined a small window of events upon cellular entry of MaBiDZ, both with (ON) and without (OFF) a magnetic field. Previous studies (25) show that nanoparticles enter cells by endocytosis, and are subsequently either stored in endosomes or lysosomes, or undergo endosomal escape. If these intracellular nanoparticles cannot escape from the endosome or lysosome, they are not available for intracellular sensing. Therefore, we investigated the distribution and colocalization of the oligo-modified MaBs and endosomes by CLSM at various time points. Results indicate that, at the peak of endosomal internalization of MaB, the ON state demonstrated about 50% less colocalization of MaB and endosomes compared to the OFF state (see SI for Figures SI7–8). Though the mechanism is under investigation, the data suggest that a magnetic field mitigates the bottleneck of endosomal sequestering, freeing nanoparticles for sensing functions in the cytoplasm.

In summary, we have designed a fluorescent hybridization MaBiDZ mRNA sensing system that can be activated by a magnetic field at the desired time. MaBiDZ sensing technology produces low backround fluorescence that can be instantly activated by magnetic field. We demonstrated that the sensor can be used for magnetic field-dependent mRNA sensing in living cells. The technology enables detection of specific mRNA in live cells within 2.5 h after applying a magnetic field, which is a significant improvement in comparison with current techniques. We hope that the MaBiDZ technology introduced here will add to the toolbox of techniques for RNA analysis in live cells. The developed approach can find much broader applications than the presently demonstrated cancer biomarker analysis example.

Supporting Information

ARTICLE SECTIONS
Jump To

The Supporting Information is available free of charge on the ACS Publications website at DOI: 10.1021/jacs.7b06022.

  • Details of experimental procedures, complete structure of probe-analyte complex, and control experiments with cell culture (PDF)

Terms & Conditions

Most electronic Supporting Information files are available without a subscription to ACS Web Editions. Such files may be downloaded by article for research use (if there is a public use license linked to the relevant article, that license may permit other uses). Permission may be obtained from ACS for other uses through requests via the RightsLink permission system: http://pubs.acs.org/page/copyright/permissions.html.

Author Information

ARTICLE SECTIONS
Jump To
  • Corresponding Authors
  • Authors
    • Saira F. Bakshi - Department of Chemistry and Biomolecular Science, Clarkson University, Potsdam, New York 13699-5810, United States
    • Nataliia Guz - Department of Chemistry and Biomolecular Science, Clarkson University, Potsdam, New York 13699-5810, United States
    • Andrey Zakharchenko - Nanostructured Materials Lab, University of Georgia, Athens, Georgia 30602, United States
    • Han Deng - Department of Biology, Clarkson University, Potsdam, New York 13699-5810, United States
    • Alexei V. Tumanov - Department of Microbiology, Immunology, and Molecular Genetics, University of Texas Health Science Center at San Antonio, 7703 Floyd Curl Drive, San Antonio, Texas 78229-3900, United States;  Orcidhttp://orcid.org/0000-0001-6042-0152
    • Craig D. Woodworth - Department of Biology, Clarkson University, Potsdam, New York 13699-5810, United States
  • Notes
    The authors declare no competing financial interest.

Acknowledgment

ARTICLE SECTIONS
Jump To

This work at Clarkson University (E.K.) and at the University of Georgia (S.M.) was supported by the NSF awards CBET-1403208 and DMR-1309469. The work at University of Central Florida (D.M.K.) and Clarkson (C.D.W.) was supported by NIH awards R15AI10388001A1 and 1R15CA173703-01. D.M.K. was partially supported by the ITMO University Fellowship and Professorship Program.

References

ARTICLE SECTIONS
Jump To

This article references 25 other publications.

  1. 1
    Germond, A.; Fujita, H.; Ichimura, T.; Watanabe, T. M. Biophys. Rev. 2016, 8, 121– 138 DOI: 10.1007/s12551-016-0195-9
    Google Scholar
    1
    Design and development of genetically encoded fluorescent sensors to monitor intracellular chemical and physical parameters
    Germond, Arno; Fujita, Hideaki; Ichimura, Taro; Watanabe, Tomonobu M.
    Biophysical Reviews (2016), 8 (2), 121-138CODEN: BRIECG; ISSN:1867-2450. (Springer)
    Over the past decades many researchers have made major contributions towards the development of genetically encoded (GE) fluorescent sensors derived from fluorescent proteins. GE sensors are now used to study biol. phenomena by facilitating the measurement of biochem. behaviors at various scales, ranging from single mols. to single cells or even whole animals. Here, we review the historical development of GE fluorescent sensors and report on their current status. We specifically focus on the development strategies of the GE sensors used for measuring pH, ion concns. (e.g., chloride and calcium), redox indicators, membrane potential, temp., pressure, and mol. crowding. We demonstrate that these fluroescent protein-based sensors have a shared history of concepts and development strategies, and we highlight the most original concepts used to date. We believe that the understanding and application of these various concepts will pave the road for the development of future GE sensors and lead to new breakthroughs in bioimaging.
    >> More from SciFinder ®
    https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC28XnsFWntrg%253D&md5=046fcb72eeb3bd731b0691767c475d8e
  2. 2
    (a) Cui, M.; You, L.; Ren, X.; Zhao, W.; Liao, Q.; Zhao, Y. Biochem. Biophys. Res. Commun. 2016, 471, 10– 14 DOI: 10.1016/j.bbrc.2015.12.101
    Google Scholar
    There is no corresponding record for this reference.
    (b) Guo, D.; Barry, L.; Lin, S. S. H.; Huang, V.; Li, L.-C. RNA Biol. 2014, 11, 1221– 1225 DOI: 10.4161/15476286.2014.972853
    Google Scholar
    2b
    RNAa in action: from the exception to the norm
    Guo Dan; Barry Liam; Lin Sharon Szu Hua; Huang Vera; Li Long-Cheng
    RNA biology (2014), 11 (10), 1221-5 ISSN:.
    Small RNA programmed Argonautes are sophisticated cellular effector platforms known to be involved in a diverse array of functions ranging from mRNA cleavage, translational inhibition, DNA elimination, epigenetic silencing, alternative splicing and even gene activation. First observed in human cells, small RNA-induced gene activation, also known as RNAa, involves the targeted recruitment of Argonaute proteins to specific promoter sequences followed by induction of stable epigenetic changes which promote transcription. The existence of RNAa remains contentious due to its elusive mechanism. A string of recent studies in C. elegans provides unequivocal evidence for RNAa's fundamental role in sculpting the epigenetic landscape and maintaining active transcription of endogenous genes and supports the presence of a functionally sophisticated network of small RNA-Argonaute pathways consisting of opposite yet complementary "yin and yang" regulatory elements. In this review, we summarize key findings from recent studies of endogenous RNAa in C. elegans, with an emphasis on the Argonaute protein CSR-1.
    >> More from SciFinder ®
    https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A280%3ADC%252BC2MvnslWnsg%253D%253D&md5=0fce92a5a3a495558ec8458e418482af
    (c) Qi, X.; Zhang, D.-H.; Wu, N.; Xiao, J.-H.; Wang, X.; Ma, W. J. Med. Genet. 2015, 52, 710– 718 DOI: 10.1136/jmedgenet-2015-103334
    Google Scholar
    2c
    ceRNA in cancer: possible functions and clinical implications
    Qi Xiaolong; Zhang Da-Hong; Wu Nan; Xiao Jun-Hua; Wang Xiang; Ma Wang
    Journal of medical genetics (2015), 52 (10), 710-8 ISSN:.
    Competing endogenous RNAs (ceRNAs) are transcripts that can regulate each other at post-transcription level by competing for shared miRNAs. CeRNA networks link the function of protein-coding mRNAs with that of non-coding RNAs such as microRNA, long non-coding RNA, pseudogenic RNA and circular RNA. Given that any transcripts harbouring miRNA response element can theoretically function as ceRNAs, they may represent a widespread form of post-transcriptional regulation of gene expression in both physiology and pathology. CeRNA activity is influenced by multiple factors such as the abundance and subcellular localisation of ceRNA components, binding affinity of miRNAs to their sponges, RNA editing, RNA secondary structures and RNA-binding proteins. Aberrations in these factors may deregulate ceRNA networks and thus lead to human diseases including cancer. In this review, we introduce the mechanisms and molecular bases of ceRNA networks, discuss their roles in the pathogenesis of cancer as well as methods of predicting and validating ceRNA interplay. At last, we discuss the limitations of current ceRNA theory, propose possible directions and envision the possibilities of ceRNAs as diagnostic biomarkers or therapeutic targets.
    >> More from SciFinder ®
    https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A280%3ADC%252BC283htV2htQ%253D%253D&md5=0489dc0a4a01909899f8c39a83130e9d
  3. 3
    Bertrand, E.; Chartrand, P.; Schaefer, M.; Shenoy, S. M.; Singer, R. H.; Long, R. M. Mol. Cell 1998, 2, 437– 445 DOI: 10.1016/S1097-2765(00)80143-4
    Google Scholar
    There is no corresponding record for this reference.
  4. 4
    Paige, J. S.; Wu, K. Y.; Jaffrey, S. R. Science 2011, 333, 642– 646 DOI: 10.1126/science.1207339
    Google Scholar
    4
    RNA mimics of green fluorescent protein
    Paige, Jeremy S.; Wu, Karen Y.; Jaffrey, Samie R.
    Science (Washington, DC, United States) (2011), 333 (6042), 642-646CODEN: SCIEAS; ISSN:0036-8075. (American Association for the Advancement of Science)
    Green fluorescent protein (GFP) and its derivs. have transformed the use and anal. of proteins for diverse applications. Like proteins, RNA has complex roles in cellular function and is increasingly used for various applications, but a comparable approach for fluorescently tagging RNA is lacking. Here, we describe the generation of RNA aptamers that bind fluorophores resembling the fluorophore in GFP. These RNA-fluorophore complexes create a palette that spans the visible spectrum. An RNA-fluorophore complex, termed Spinach, resembles enhanced GFP and emits a green fluorescence comparable in brightness with fluorescent proteins. Spinach is markedly resistant to photobleaching, and Spinach fusion RNAs can be imaged in living cells. These RNA mimics of GFP provide an approach for genetic encoding of fluorescent RNAs.
    >> More from SciFinder ®
    https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3MXpt1yisbw%253D&md5=dc757f578e744c78062a68b52d15a961
  5. 5
    (a) Holtzer, L.; Oleinich, I.; Anzola, M.; Lindberg, E.; Sadhu, K. K.; Gonzalez-Gaitan, M.; Winssinger, N. ACS Cent. Sci. 2016, 2, 394– 400 DOI: 10.1021/acscentsci.6b00054
    Google Scholar
    5a
    Nucleic Acid Templated Chemical Reaction in a Live Vertebrate
    Holtzer, Laurent; Oleinich, Igor; Anzola, Marcello; Lindberg, Eric; Sadhu, Kalyan K.; Gonzalez-Gaitan, Marcos; Winssinger, Nicolas
    ACS Central Science (2016), 2 (6), 394-400CODEN: ACSCII; ISSN:2374-7951. (American Chemical Society)
    Nucleic acid templated reactions are enabled by the hybridization of probe-reagent conjugates resulting in high effective reagent concn. and fast chem. transformation. We have developed a reaction that harnesses cellular microRNA (miRNA) to yield the cleavage of a linker releasing fluorogenic rhodamine in a live vertebrate. The reaction is based on the catalytic photoredn. of an azide by a ruthenium complex. We showed that this system reports specific expression of miRNA in living tissues of a vertebrate.
    >> More from SciFinder ®
    https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC28XotFelsLo%253D&md5=241e3e5dcd20cdf43600dd27137b9111
    (b) Michaelis, J.; Roloff, A.; Seitz, O. Org. Biomol. Chem. 2014, 12, 2821– 2833 DOI: 10.1039/C4OB00096J
    Google Scholar
    There is no corresponding record for this reference.
    (c) Franzini, R. M.; Kool, E. T. Bioconjugate Chem. 2011, 22, 1869– 1877 DOI: 10.1021/bc2003567
    Google Scholar
    5c
    Improved Templated Fluorogenic Probes Enhance the Analysis of Closely Related Pathogenic Bacteria by Microscopy and Flow Cytometry
    Franzini, Raphael M.; Kool, Eric T.
    Bioconjugate Chemistry (2011), 22 (9), 1869-1877CODEN: BCCHES; ISSN:1043-1802. (American Chemical Society)
    Templated fluorescence activation has recently emerged as a promising mol. approach to detect and differentiate nucleic acid sequences in vitro and in cells. Here, we describe the application of a reductive quencher release strategy to the taxonomic anal. of Gram-neg. bacteria by targeting a single nucleotide difference in their 16S rRNA in a two-color assay. For this purpose, it was necessary to develop a release linker contg. a quencher suitable for red and near-IR fluorophores, and to improve methods for the delivery of probes into cells. A cyanine-dye labeled oligonucleotide probe contg. the new quencher-release linker showed unprecedentedly low background signal and high fluorescence turn-on ratios. The combination of a fluorescein-contg. and a near-IR emitting probe discriminated E. coli from S. enterica despite nearly identical ribosomal target sequences. Two-color anal. by microscopy and the first successful discrimination of bacteria by two-color flow cytometry with templated reactive probes are described.
    >> More from SciFinder ®
    https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3MXhtVOktrbK&md5=92ba463514226e5eda1ed0cdf39be94f
  6. 6
    Santangelo, P. J.; Nix, B.; Tsourkas, A.; Bao, G. Nucleic Acids Res. 2004, 32, e57 DOI: 10.1093/nar/gnh062
    Google Scholar
    6
    Dual FRET molecular beacons for mRNA detection in living cells
    Santangelo, Philip J.; Nix, Brent; Tsourkas, Andrew; Bao, Gang
    Nucleic Acids Research (2004), 32 (6), e57/1-e57/9CODEN: NARHAD; ISSN:0305-1048. (Oxford University Press)
    The ability to visualize in real-time the expression level and localization of specific endogenous RNAs in living cells can offer tremendous opportunities for biol. and disease studies. Here we demonstrate such a capability using a pair of mol. beacons, one with a donor and the other with an acceptor fluorophore that hybridize to adjacent regions on the same mRNA target, resulting in fluorescence resonance energy transfer (FRET). Detection of the FRET signal significantly reduced false positives, leading to sensitive imaging of K-ras and survivin mRNAs in live HDF and MIAPaCa-2 cells. FRET detection gave a ratio of 2.25 of K-ras mRNA expression in stimulated and unstimulated HDF, comparable to the ratio of 1.95 using RT-PCR, and in contrast to the single-beacon result of 1.2. We further revealed intriguing details of K-ras and survivin mRNA localization in living cells. The dual FRET mol. beacons approach provides a novel technique for sensitive RNA detection and quantification in living cells.
    >> More from SciFinder ®
    https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD2cXjt1ejsb8%253D&md5=3a808296da2f8bc0ad8f5b6bc20e5f7f
  7. 7
    (a) Zhao, D.; Yang, Y.; Qu, N.; Chen, M.; Ma, Z.; Krueger, C. J.; Behlke, M. A.; Chen, A. K. Biomaterials 2016, 100, 172– 183 DOI: 10.1016/j.biomaterials.2016.05.022
    Google Scholar
    7a
    Single-molecule detection and tracking of RNA transcripts in living cells using phosphorothioate-optimized 2'-O-methyl RNA molecular beacons
    Zhao, Dan; Yang, Yantao; Qu, Na; Chen, Mingming; Ma, Zhao; Krueger, Christopher J.; Behlke, Mark A.; Chen, Antony K.
    Biomaterials (2016), 100 (), 172-183CODEN: BIMADU; ISSN:0142-9612. (Elsevier Ltd.)
    Mol. beacons (MBs) composed of 2'-O-Me RNA (2Me) and phosphorothioate (PS) linkages throughout the backbone (2Me/PSFULL MBs) have enabled long-term imaging of RNA in living cells, but excess PS modification can induce nonspecific binding, causing false-pos. signals. In this study, we evaluate the intracellular stability of MBs composed of 2Me with various PS modifications, and found that false-pos. signals could be reduced to marginal levels when the MBs possess a fully PS-modified loop domain and a phosphodiester stem (2Me/PSLOOP MB). Addnl., 2Me/PSLOOP MBs exhibited uncompromised hybridization kinetics, prolonged functionality and >88% detection accuracy for single RNA transcripts, and could do so without interfering with gene expression or cell growth. Finally, 2Me/PSLOOP MBs could image the dynamics of single mRNA transcripts in the nucleus and the cytoplasm simultaneously, regardless of whether the MBs targeted the 5'- or the 3'-UTR. Together, these findings demonstrate the effectiveness of loop-domain PS modification in reducing nonspecific signals and the potential for sensitive and accurate imaging of individual RNAs at the single-mol. level. With the growing interest in the role of RNA localization and dynamics in health and disease, 2Me/PSLOOP MBs could enable new discoveries in RNA research.
    >> More from SciFinder ®
    https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC28Xos12rtbk%253D&md5=1c70b0aa6afd761532be7911f168601d
    (b) Giraldo-Vela, J. P.; Kang, W.; McNaughton, R. L.; Zhang, X.; Wile, B. M.; Tsourkas, A.; Bao, G.; Espinosa, H. D. Small 2015, 11, 2386– 2391 DOI: 10.1002/smll.201401137
    Google Scholar
    There is no corresponding record for this reference.
  8. 8
    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
    8
    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.
    >> More from SciFinder ®
    https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD2sXhtlKru7jE&md5=71e766b235655781c09ccfc331b37762
    (a) Prigodich, A. E.; Seferos, D. S.; Massich, M. D.; Giljohann, D. A.; Lane, B. C.; Mirkin, C. A. ACS Nano 2009, 3, 2147– 2152 DOI: 10.1021/nn9003814
    Google Scholar
    There is no corresponding record for this reference.
    (b) Zheng, D.; Seferos, D. S.; Giljohann, D. A.; Patel, P. C.; Mirkin, C. A. Nano Lett. 2009, 9, 3258– 3261 DOI: 10.1021/nl901517b
    Google Scholar
    8b
    Aptamer Nano-flares for Molecular Detection in Living Cells
    Zheng, Dan; Seferos, Dwight S.; Giljohann, David A.; Patel, Pinal C.; Mirkin, Chad A.
    Nano Letters (2009), 9 (9), 3258-3261CODEN: NALEFD; ISSN:1530-6984. (American Chemical Society)
    The authors demonstrate a composite nanomaterial, termed an aptamer nano-flare, that can directly quantify an intracellular analyte in a living cell. Aptamer nano-flares consist of a gold nanoparticle core functionalized with a dense monolayer of nucleic acid aptamers with a high affinity for ATP. The probes bind selectively to target mols. and release fluorescent reporters which indicate the presence of the analyte. Addnl., these nanoconjugates are readily taken up by cells where their signal intensity can be used to quantify intracellular analyte concn. These nanoconjugates are a promising approach for the intracellular quantification of other small mols. or proteins, or as agents that use aptamer binding to elicit a biol. response in living systems.
    >> More from SciFinder ®
    https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD1MXptlSmtbw%253D&md5=ed0a131b9ccca7002b24570f3032628b
    (c) Pan, W.; Li, Y.; Wang, M.; Yang, H.; Li, N.; Tang, B. Chem. Commun. 2016, 52, 4569– 4572 DOI: 10.1039/C5CC10147F
    Google Scholar
    There is no corresponding record for this reference.
  9. 9
    Briley, W. E.; Bondy, M. H.; Randeria, P. S.; Dupper, T. J.; Mirkin, C. A. Proc. Natl. Acad. Sci. U. S. A. 2015, 112, 9591– 9595 DOI: 10.1073/pnas.1510581112
    Google Scholar
    There is no corresponding record for this reference.
  10. 10
    (a) Feyder, M.; Goff, L. A. J. Clin. Invest. 2016, 126, 2783– 2791 DOI: 10.1172/JCI84422
    Google Scholar
    There is no corresponding record for this reference.
    (b) Ouellet, J. Front. Chem. 2016, 429 DOI: 10.3389/fchem.2016.00029
    Google Scholar
    10b
    RNA Fluorescence with Light-Up Aptamers
    Ouellet Jonathan
    Frontiers in chemistry (2016), 4 (), 29 ISSN:2296-2646.
    Seeing is not only believing; it also includes understanding. Cellular imaging with GFP in live cells has been transformative in many research fields. Modulation of cellular regulation is tightly regulated and innovative imaging technologies contribute to further understand cellular signaling and physiology. New types of genetically encoded biosensors have been developed over the last decade. They are RNA aptamers that bind with their cognate fluorogen ligands and activate their fluorescence. The emergence and the evolution of these RNA aptamers as well as their conversion into a wide spectrum of applications are examined in a global way.
    >> More from SciFinder ®
    https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A280%3ADC%252BC2s3jsFanug%253D%253D&md5=ebf0cfee8a0de42cbef6c299822eb033
    (c) Shigeto, H.; Nakatsuka, K.; Ikeda, T.; Hirota, R.; Kuroda, A.; Funabashi, H. Anal. Chem. 2016, 88, 7894– 7898 DOI: 10.1021/acs.analchem.6b02710
    Google Scholar
    There is no corresponding record for this reference.
    (d) Takahashi, K.; Ito, S.; Nakamoto, K.; Ito, Y.; Ueno, Y. J. Org. Chem. 2015, 80, 8561– 8570 DOI: 10.1021/acs.joc.5b01132
    Google Scholar
    There is no corresponding record for this reference.
    (e) Urbanek, M. O.; Galka-Marciniak, P.; Olejniczak, M.; Krzyzosiak, W. J. RNA Biol. 2014, 11, 1083– 1095 DOI: 10.4161/rna.35506
    Google Scholar
    10e
    RNA imaging in living cells - methods and applications
    Urbanek Martyna O; Galka-Marciniak Paulina; Olejniczak Marta; Krzyzosiak Wlodzimierz J
    RNA biology (2014), 11 (8), 1083-95 ISSN:.
    Numerous types of transcripts perform multiple functions in cells, and these functions are mainly facilitated by the interactions of the RNA with various proteins and other RNAs. Insight into the dynamics of RNA biosynthesis, processing and cellular activities is highly desirable because this knowledge will deepen our understanding of cell physiology and help explain the mechanisms of RNA-mediated pathologies. In this review, we discuss the live RNA imaging systems that have been developed to date. We highlight information on the design of these systems, briefly discuss their advantages and limitations and provide examples of their numerous applications in various organisms and cell types. We present a detailed examination of one application of RNA imaging systems: this application aims to explain the role of mutant transcripts in human disease pathogenesis caused by triplet repeat expansions. Thus, this review introduces live RNA imaging systems and provides a glimpse into their various applications.
    >> More from SciFinder ®
    https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A280%3ADC%252BC2MzmtVSgsg%253D%253D&md5=00375391046f0df6d39ca4dc084e0eeb
  11. 11
    (a) Wang, C.; Zhu, Z.; Song, Y.; Lin, H.; Yang, C. J.; Tan, W. Chem. Commun. 2011, 47, 5708– 5710 DOI: 10.1039/c1cc10481k
    Google Scholar
    There is no corresponding record for this reference.
    (b) Joshi, K. B.; Vlachos, A.; Mikat, V.; Deller, T.; Heckel, A. Chem. Commun. 2012, 48, 2746– 2748 DOI: 10.1039/C2CC16654B
    Google Scholar
    There is no corresponding record for this reference.
    (c) Rinne, J. S.; Kaminski, T. P.; Kubitscheck, U.; Heckel, A. Chem. Commun. 2013, 49, 5375– 5377 DOI: 10.1039/c3cc42420k
    Google Scholar
    There is no corresponding record for this reference.
  12. 12
    (a) Ruble, B. K.; Yeldell, S. B.; Dmochowski, I. J. J. Inorg. Biochem. 2015, 150, 182– 188 DOI: 10.1016/j.jinorgbio.2015.03.010
    Google Scholar
    There is no corresponding record for this reference.
    (b) Tang, X.; Zhang, J.; Sun, J.; Wang, Y.; Wu, J.; Zhang, L. Org. Biomol. Chem. 2013, 11, 7814– 7824 DOI: 10.1039/c3ob41735b
    Google Scholar
    There is no corresponding record for this reference.
    (c) Bort, G.; Gallavardin, T.; Ogden, D.; Dalko, P. I. Angew. Chem., Int. Ed. 2013, 52, 4526– 4537 DOI: 10.1002/anie.201204203
    Google Scholar
    12c
    From One-Photon to Two-Photon Probes: "Caged" Compounds, Actuators, and Photoswitches
    Bort, Guillaume; Gallavardin, Thibault; Ogden, David; Dalko, Peter I.
    Angewandte Chemie, International Edition (2013), 52 (17), 4526-4537CODEN: ACIEF5; ISSN:1433-7851. (Wiley-VCH Verlag GmbH & Co. KGaA)
    A review. Mol. systems that can be remotely controlled by light are gaining increasing importance in cell biol., physiol., and neurosciences because of the spatial and temporal precision that is achievable with laser microscopy. Two-photon excitation has significant advantages deep in biol. tissues, but raises problems in the design of "smart" probes compatible with cell physiol. This Review discusses the chem. challenges in generating suitable two-photon probes.
    >> More from SciFinder ®
    https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3sXislWgsb4%253D&md5=0d8affb2b63341dcbad735299c2464ec
  13. 13
    (a) Luo, D.; Saltzman, W. M. Nat. Biotechnol. 2000, 18, 33– 37 DOI: 10.1038/71889
    Google Scholar
    There is no corresponding record for this reference.
    (b) Patil, S. D.; Rhodes, D. G.; Burgess, D. J. AAPS J. 2005, 7, E61– 77 DOI: 10.1208/aapsj070109
    Google Scholar
    There is no corresponding record for this reference.
    (c) Bishop, C. J.; Kozielski, K. L.; Green, J. J. J. Controlled Release 2015, 219, 488– 499 DOI: 10.1016/j.jconrel.2015.09.046
    Google Scholar
    There is no corresponding record for this reference.
  14. 14
    Dobson, J. Nat. Nanotechnol. 2008, 3, 139– 143 DOI: 10.1038/nnano.2008.39
    Google Scholar
    14
    Remote control of cellular behaviour with magnetic nanoparticles
    Dobson, Jon
    Nature Nanotechnology (2008), 3 (3), 139-143CODEN: NNAABX; ISSN:1748-3387. (Nature Publishing Group)
    A review. By binding magnetic nanoparticles to the surface of cells, it is possible to manipulate and control cell function with an external magnetic field. The technique of activating cells with magnetic nanoparticles offers a means to isolate and explore cellular mechanics and ion channel activation to gain better understanding of these processes. Here, we go beyond using this technique as an investigative tool and focus on its potential to actively control cellular functions and processes with an eye towards biol. and clin. applications. In particular, we focus on applications in tissue engineering and regenerative medicine.
    >> More from SciFinder ®
    https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD1cXivFKgtrY%253D&md5=700c579988781da1d25c69e9023eb705
  15. 15
    (a) Wang, N.; Butler, J. P.; Ingber, D. E. Science 1993, 260, 1124– 1127 DOI: 10.1126/science.7684161
    Google Scholar
    There is no corresponding record for this reference.
    (b) Meyer, C. J.; Alenghat, F. J.; Rim, P.; Fong, J. H.-J.; Fabry, B.; Ingber, D. E. Nat. Cell Biol. 2000, 2, 666– 668 DOI: 10.1038/35023621
    Google Scholar
    15b
    Mechanical control of cyclic AMP signalling and gene transcription through integrins
    Meyer, Christian J.; Alenghat, Francis J.; Rim, Paul; Fong, Jenny Hwai-Jen; Fabry, Ben; Ingber, Donald E.
    Nature Cell Biology (2000), 2 (9), 666-668CODEN: NCBIFN; ISSN:1465-7392. (Nature Publishing Group)
    This study was carried out to discriminate between two alternative hypotheses as to how cells sense mech. forces and transduce them into changes in gene transcription. Do cells sense mech. signals through generalized membrane distortion or through specific transmembrane receptors, such as integrins. Here the authors show that mech. stresses applied to the cell surface alter the cAMP signalling cascade and downstream gene transcription by modulating local release of signals generated by activated integrin receptors in a G-protein-dependent manner, whereas distortion of integrins in the absence of receptor occupancy has no effect.
    >> More from SciFinder ®
    https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD3cXmslGks7k%253D&md5=033770918a88216173175fb8aaec4e52
  16. 16
    (a) Bausch, A. R.; Hellerer, U.; Essler, M.; Aepfelbacher, M.; Sackmann, E. Biophys. J. 2001, 80, 2649– 2657 DOI: 10.1016/S0006-3495(01)76234-0
    Google Scholar
    There is no corresponding record for this reference.
    (b) Bausch, A. R.; Möller, W.; Sackmann, E. Biophys. J. 1999, 76, 573– 579 DOI: 10.1016/S0006-3495(99)77225-5
    Google Scholar
    16b
    Measurement of local viscoelasticity and forces in living cells by magnetic tweezers
    Bausch, Andreas R.; Moller, Winfried; Sackmann, Erich
    Biophysical Journal (1999), 76 (1, Pt. 1), 573-579CODEN: BIOJAU; ISSN:0006-3495. (Biophysical Society)
    We measured the viscoelastic properties of the cytoplasm of J774 macrophages with a recently developed microrheometer. Ferromagnetic beads (1.3 μm in diam.) were used to det. the local viscoelastic moduli. Step-force pulses were applied to the magnetic beads and the displacement was obsd. by single particle tracking. By analyzing the creep response curves in terms of a triphasic mech. equiv. circuit, we measured the shear elastic modulus, the effective viscosities, and the strain relaxation time. The values of the shear modulus vary by more than an order of magnitude within the cell population (range, 20-735 Pa; av., 343 Pa) and by a factor of 2 within single cells. The effective viscosity of the cytoplasm exhibits a relatively sharp distribution about an av. of η = 210 Pa s (± 143 Pa s). We measured the displacement field generated by the local forces by observing the induced motion of nonmagnetic beads. Even at distances of the order of 1 μm, no induced motion was seen, suggesting that the cytoplasm is composed of clusters of densely packed and cross-linked filaments sepd. by soft regions. In another series of expts. we analyzed the magnetophoretic motion of the ferromagnetic beads at a const. magnetic force. Measuring the bead velocity parallel and perpendicular to the applied force showed that local active forces on the beads varied from 50 to 900 pN.
    >> More from SciFinder ®
    https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADyaK1MXjs1Sjug%253D%253D&md5=c7cfba90639225899ff9ac732ecdc983
    (c) Glogauer, M.; Ferrier, J. Pfluegers Arch. 1997, 435, 320– 327 DOI: 10.1007/s004240050518
    Google Scholar
    There is no corresponding record for this reference.
  17. 17
    Hughes, S.; McBain, S.; Dobson, J.; El Haj, A. J. J. J. R. Soc., Interface 2008, 5, 855– 863 DOI: 10.1098/rsif.2007.1274
    Google Scholar
    17
    Selective activation of mechanosensitive ion channels using magnetic particles
    Hughes, Steven; McBain, Stuart; Dobson, Jon; El Haj, Alicia J.
    Journal of the Royal Society, Interface (2008), 5 (25), 855-863CODEN: JRSICU; ISSN:1742-5689. (Royal Society)
    This study reports the preliminary development of a novel magnetic particle-based technique that permits the application of highly localized mech. forces directly to specific regions of an ion-channel structure. We demonstrate that this approach can be used to directly and selectively activate a mechanosensitive ion channel of interest, namely TREK-1. It is shown that manipulation of particles targeted against the extended extracellular loop region of TREK-1 leads to changes in whole-cell currents consistent with changes in TREK-1 activity. Responses were absent when particles were coated with RGD (Arg-Gly-Asp) peptide or when magnetic fields were applied in the absence of magnetic particles. It is concluded that changes in whole-cell current are the result of direct force application to the extracellular loop mechano-gating. It is hypothesized that the extended loop region of TREK-1 may act as a tension spring that acts to regulate sensitivity to mech. forces, in a nature similar to that described for MscL. The development of a technique that permits the direct manipulation of mechanosensitive ion channels in real time without the need for pharmacol. drugs has huge potential benefits not only for basic biol. research of ion-channel gating mechanisms, but also potentially as a tool for the treatment of human diseases caused by ion-channel dysfunction.
    >> More from SciFinder ®
    https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD1cXpvVGntLc%253D&md5=a114c2e9f738827ec18e4e5c25a68f75
  18. 18
    (a) Kolpashchikov, D. M. ChemBioChem 2007, 8, 2039– 2042 DOI: 10.1002/cbic.200700384
    Google Scholar
    There is no corresponding record for this reference.
    (b) Mokany, E.; Bone, S. M.; Young, P. E.; Doan, T. B.; Todd, A. V. J. Am. Chem. Soc. 2010, 132, 1051– 1059 DOI: 10.1021/ja9076777
    Google Scholar
    18b
    MNAzymes, a Versatile New Class of Nucleic Acid Enzymes That Can Function as Biosensors and Molecular Switches
    Mokany, Elisa; Bone, Simon M.; Young, Paul E.; Doan, Tram B.; Todd, Alison V.
    Journal of the American Chemical Society (2010), 132 (3), 1051-1059CODEN: JACSAT; ISSN:0002-7863. (American Chemical Society)
    To increase the versatility and utility of nucleic acid enzymes, multicomponent complexes known as MNAzymes were developed, which produce amplified output signals in response to specific input signals. Multiple oligonucleotide partzymes assemble into active MNAzymes only in the presence of an input assembly facilitator such as a target nucleic acid. Once formed, MNAzymes catalytically modify a generic substrate, generating an amplified output signal that heralds the presence of the target while leaving the target intact. Several applications were demonstrated, including sensitive, isothermal target detection; discrimination of polymorphisms; and highly specific monitoring of real-time polymerase chain reaction (PCR). Furthermore, their capacity to function as mol. switches and to work in series to create a mol. cascade was demonstrated. The modular nature of MNAzymes, together with the sepn. of input and output functionalities, provides potential for their integration into diverse devices such as diagnostic biosensors, mol. computers, and/or nanoscale machines.
    >> More from SciFinder ®
    https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD1MXhs1WjsLvF&md5=80a44a8993aa11039a48e664af73d761
    (c) Gerasimova, Y. V.; Cornett, E.; Kolpashchikov, D. M. ChemBioChem 2010, 11, 811– 817 DOI: 10.1002/cbic.201000006
    Google Scholar
    There is no corresponding record for this reference.
    (d) Gerasimova, Y. V.; Kolpashchikov, D. M. Angew. Chem., Int. Ed. 2013, 52, 10586– 10588 DOI: 10.1002/anie.201303919
    Google Scholar
    There is no corresponding record for this reference.
  19. 19
    Bumb, A.; Brechbiel, M. W.; Choyke, P. L.; Fugger, L.; Eggeman, A.; Prabhakaran, D.; Hutchinson, J.; Dobson, P. J. Nanotechnology 2008, 19335601 DOI: 10.1088/0957-4484/19/33/335601
    Google Scholar
    There is no corresponding record for this reference.
  20. 20
    (a) Gref, R.; Lück, M.; Quellec, P.; Marchand, M.; Dellacherie, E.; Harnisch, S.; Blunk, T.; Müller, R. H. Colloids Surf., B 2000, 18, 301– 313 DOI: 10.1016/S0927-7765(99)00156-3
    Google Scholar
    There is no corresponding record for this reference.
  21. 21
    Cheng, G. Z.; Chan, J.; Wang, Q.; Zhang, W. Z.; Sun, C. D.; Wang, L. H. Cancer Res. 2007, 67, 1979– 1987 DOI: 10.1158/0008-5472.CAN-06-1479
    Google Scholar
    There is no corresponding record for this reference.
  22. 22
    Watanabe, O.; Imamura, H.; Shimizu, T.; Kinoshita, J.; Okabe, T.; Hirano, A.; Yoshimatsu, K.; Konno, S.; Aiba, M.; Ogawa, K. Anticancer Res. 2004, 24, 3851– 3856
    Google Scholar
    22
    Expression of Twist and Wnt in human breast cancer
    Watanabe, Osamu; Imamura, Hiroshi; Shimizu, Tadao; Kinoshita, Jun; Okabe, Toshihiro; Hirano, Akira; Yoshimatsu, Kazuhiko; Konno, Soichi; Aiba, Motohiko; Ogawa, Kenji
    Anticancer Research (2004), 24 (6), 3851-3856CODEN: ANTRD4; ISSN:0250-7005. (International Institute of Anticancer Research)
    Twist, a transcription factor of the basic helix-loop-helix class, has been suggested to have oncogenic properties. We reported Twist expression was regulated by Wnt/β-catenin signaling and that both Wnt-1 and Twist could contribute to mammary tumorigenesis. The aim of this study was to demonstrate the expression of Twist, Wnt-1 and Wnt-2 in human breast cancer tissue. We examd. the expression in patients with breast cancer by RT-PCR and immunohistochem. RT-PCR of twenty-three pairs of cancer and normal breast tissue revealed that Twist was up-regulated in 69.6% (16/23) of the cancer lesions and 21.7% (5/23) of the normal breast tissues. Wnt-2 was up-regulated in all of the cancer lesions and 13.0% (3/23) of the normal breast tissues, whereas Wnt-1 was expressed in both the cancer and normal breast tissues of the five cases examd. Immunohistochem. analyses revealed that Twist was pos. expressed in 52.2% (12/23) of the cancer lesions and 34.8% (8/23) of the normal breast tissues. Twist and Wnt-2 are highly expressed in breast cancer tissue, suggesting that both mols. could play important roles in mammary carcinogenesis.
    >> More from SciFinder ®
    https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD2MXitFylsLk%253D&md5=23684de587fa14fb044471b29d846ab0
  23. 23
    Li, Y.; Wang, W.; Wang, W.; Yang, R.; Wang, T.; Su, T.; Weng, D.; Tao, T.; Li, W.; Ma, D.; Wang, S. Gynecol. Oncol. 2012, 124, 112– 118 DOI: 10.1016/j.ygyno.2011.09.003
    Google Scholar
    23
    Correlation of TWIST2 up-regulation and epithelial-mesenchymal transition during tumorigenesis and progression of cervical carcinoma
    Li, Yan; Wang, Wei; Wang, Wenwen; Yang, Runfeng; Wang, Tian; Su, Tiefen; Weng, Danhui; Tao, Tao; Li, Wei; Ma, Ding; Wang, Shixuan
    Gynecologic Oncology (2012), 124 (1), 112-118CODEN: GYNOA3; ISSN:0090-8258. (Elsevier B.V.)
    Globally, cervical cancer is the second most common cancer among women, and detg. potential targets involved in tumor progression is necessary. This study investigated the clinic-pathol. significance of twist homolog 2 (TWIST2), a basic helix-loop-helix transcription factor, and correlated TWIST2 and E-cadherin expression in cervical cancer. A series of 142 samples, including 14 cases of normal cervical tissues, 58 cases of cervical intraepithelial neoplasia (CIN) and 70 cases of squamous cell carcinoma (SCC), were examd. TWIST2 and E-cadherin immunohistochem. staining and statistical anal. Increased cytoplasmic and nuclear expression levels of TWIST2 were assocd. with the malignant transformation of cervical epithelium and the histol. progression of cervical cancer. A logistic test showed that TWIST2 was a relatively independent predictor of lymph node metastasis of SCC. Further, increased levels of TWIST2 were also assocd. with aberrant expression of E-cadherin, an important EMT indicator. The present data suggest that TWIST2 overexpression was significantly linked to cervical cancer progression, which makes it a promising marker for detg. the metastatic potential of cervical cancer, and up-regulation of TWIST2, in combination with aberrant E-cadherin expression in primary cervical cancer tissues, may predict the malignant transformation and distal metastasis of carcinomas.
    >> More from SciFinder ®
    https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3MXhsF2rtLjL&md5=8258c2465a996782d558c320554942aa
  24. 24
    Plank, C.; Schillinger, U.; Scherer, F.; Bergemann, C.; Rémy, J.-S.; Krötz, F.; Anton, M.; Lausier, J.; Rosenecker, J. Biol. Chem. 2003, 384, 737– 747 DOI: 10.1515/BC.2003.082
    Google Scholar
    There is no corresponding record for this reference.
  25. 25
    Nguyen, J.; Szoka, F. Acc. Chem. Res. 2012, 45, 1153– 1162 DOI: 10.1021/ar3000162
    Google Scholar
    There is no corresponding record for this reference.

Cited By

ARTICLE SECTIONS
Jump To

This article is cited by 43 publications.

  1. Yiren Cao, Hongquan Zhang, X. Chris Le. Split Locations and Secondary Structures of a DNAzyme Critical to Binding-Assembled Multicomponent Nucleic Acid Enzymes for Protein Detection. Analytical Chemistry 2021, 93 (47) , 15712-15719. https://doi.org/10.1021/acs.analchem.1c03617
  2. Cheng Fang, Yuming Li, Song Hu, Hao Wang, Xiaoxia Chen, Xiaoli Zhu. Self-Assembled Growing DNA Tree Mediated by Exosomes for Amplified Imaging of Messenger RNA in Living Cells. Analytical Chemistry 2021, 93 (24) , 8414-8422. https://doi.org/10.1021/acs.analchem.1c00211
  3. Qi Kang, Man He, Beibei Chen, Guangyang Xiao, Bin Hu. MNAzyme-Catalyzed Amplification Assay with Lanthanide Tags for the Simultaneous Detection of Multiple microRNAs by Inductively Coupled Plasma–Mass Spectrometry. Analytical Chemistry 2021, 93 (2) , 737-744. https://doi.org/10.1021/acs.analchem.0c02455
  4. Rongxing Zhou, Changjia Hu, Yanwen Jin, Jie Zhang, Huan Du, Peng Yang, Junbo Chen, Xiandeng Hou, Nansheng Cheng. Spatially Constrained DNA Nanomachines To Accelerate Kinetics in Response to External Input: Design and Bioanalysis. Analytical Chemistry 2020, 92 (13) , 8909-8916. https://doi.org/10.1021/acs.analchem.0c00802
  5. Jian Zhao, Hongqian Chu, Ya Zhao, Yi Lu, Lele Li. A NIR Light Gated DNA Nanodevice for Spatiotemporally Controlled Imaging of MicroRNA in Cells and Animals. Journal of the American Chemical Society 2019, 141 (17) , 7056-7062. https://doi.org/10.1021/jacs.9b01931
  6. Meihua Lin, Xiaoqing Yi, Fujian Huang, Xin Ma, Xiaolei Zuo, Fan Xia. Photoactivated Nanoflares for mRNA Detection in Single Living Cells. Analytical Chemistry 2019, 91 (3) , 2021-2027. https://doi.org/10.1021/acs.analchem.8b04434
  7. Yudie Sun, Tao Li. Composition-Tunable Hollow Au/Ag SERS Nanoprobes Coupled with Target-Catalyzed Hairpin Assembly for Triple-Amplification Detection of miRNA. Analytical Chemistry 2018, 90 (19) , 11614-11621. https://doi.org/10.1021/acs.analchem.8b03067
  8. Bo Tian, Yuanyuan Han, Erik Wetterskog, Marco Donolato, Mikkel Fougt Hansen, Peter Svedlindh, Mattias Strömberg. MicroRNA Detection through DNAzyme-Mediated Disintegration of Magnetic Nanoparticle Assemblies. ACS Sensors 2018, 3 (9) , 1884-1891. https://doi.org/10.1021/acssensors.8b00850
  9. Fujian Huang, Meihua Lin, Ruilin Duan, Xiaoding Lou, Fan Xia, Itamar Willner. Photoactivated Specific mRNA Detection in Single Living Cells by Coupling “Signal-on” Fluorescence and “Signal-off” Electrochemical Signals. Nano Letters 2018, 18 (8) , 5116-5123. https://doi.org/10.1021/acs.nanolett.8b02004
  10. Xin Li, Yue Gao, Helin Li, Jean-Pierre Majoral, Xiangyang Shi, Andrij Pich. Smart and bioinspired systems for overcoming biological barriers and enhancing disease theranostics. Progress in Materials Science 2023, 140 , 101170. https://doi.org/10.1016/j.pmatsci.2023.101170
  11. Wei Xu, Feifei Hu, Jiajing Li, Jinhua Shang, Xiaoqing Liu, Yan Zeng, Qiong Wu, Fuan Wang. External stimulation-controlled dynamic DNA devices for biosensing and biomedical applications. Science China Chemistry 2023, 66 (11) , 3105-3115. https://doi.org/10.1007/s11426-023-1783-5
  12. Qiong Wu, Mengqing Xu, Jinhua Shang, Shizhen He, Xiaoqing Liu, Fuan Wang. Stimuli‐Responsive DNA Circuits for High‐Performance Bioimaging Application. Advanced Sensor Research 2023, 2 (9) https://doi.org/10.1002/adsr.202200102
  13. Jian Wang, Wenjie Zhang, Yating Zhang, Haolin Li. Preparation of Polymer-Based Nano-Assembled Particles with Fe3O4 in the Core. Polymers 2023, 15 (11) , 2498. https://doi.org/10.3390/polym15112498
  14. M. Mustafa Azeem, Muhammad Shafa, Muhammad Aamir, Muhammad Zubair, Basma Souayeh, Mir Waqas Alam. Nucleotide detection mechanism and comparison based on low-dimensional materials: A review. Frontiers in Bioengineering and Biotechnology 2023, 11 https://doi.org/10.3389/fbioe.2023.1117871
  15. Qing Wang, Zeyue Wang, Yuqiu He, Bin Xiong, Yingfu Li, Fuan Wang. Chemical and structural modification of RNA-cleaving DNAzymes for efficient biosensing and biomedical applications. TrAC Trends in Analytical Chemistry 2023, 159 , 116910. https://doi.org/10.1016/j.trac.2022.116910
  16. Shivkanya Fuloria, Vetriselvan Subramaniyan, Gaurav Gupta, Mahendran Sekar, Dhanalekshmi Unnikrishnan Meenakshi, Kathiresan Sathasivam, Kalvatala Sudhakar, Khalid Saad Alharbi, Sultan Saadi Almutairi, Waleed Hassan Almalki, Neeraj Kumar Fuloria. Detection of Circulating Tumor Cells and Epithelial Progenitor Cells: A Comprehensive Study. Journal of Environmental Pathology, Toxicology and Oncology 2023, 42 (3) , 1-29. https://doi.org/10.1615/JEnvironPatholToxicolOncol.2022044456
  17. Aya Shibata, Sayuri L. Higashi, Masato Ikeda. Nucleic acid-based fluorescent sensor systems: a review. Polymer Journal 2022, 54 (6) , 751-766. https://doi.org/10.1038/s41428-022-00623-1
  18. Zhichu Xiang, Jian Zhao, Deyu Yi, Zhenghan Di, Lele Li. Peptide Nucleic Acid (PNA)‐Guided Peptide Engineering of an Aptamer Sensor for Protease‐Triggered Molecular Imaging. Angewandte Chemie 2021, 133 (42) , 22841-22845. https://doi.org/10.1002/ange.202106639
  19. Zhichu Xiang, Jian Zhao, Deyu Yi, Zhenghan Di, Lele Li. Peptide Nucleic Acid (PNA)‐Guided Peptide Engineering of an Aptamer Sensor for Protease‐Triggered Molecular Imaging. Angewandte Chemie International Edition 2021, 60 (42) , 22659-22663. https://doi.org/10.1002/anie.202106639
  20. Erin M. McConnell, Ioana Cozma, Quanbing Mou, John D. Brennan, Yi Lu, Yingfu Li. Biosensing with DNAzymes. Chemical Society Reviews 2021, 50 (16) , 8954-8994. https://doi.org/10.1039/D1CS00240F
  21. Ioana Cozma, Erin M. McConnell, John D. Brennan, Yingfu Li. DNAzymes as key components of biosensing systems for the detection of biological targets. Biosensors and Bioelectronics 2021, 177 , 112972. https://doi.org/10.1016/j.bios.2021.112972
  22. Xue Gong, Ruomeng Li, Jing Wang, Jie Wei, Kang Ma, Xiaoqing Liu, Fuan Wang. A Smart Theranostic Nanocapsule for Spatiotemporally Programmable Photo‐Gene Therapy. Angewandte Chemie 2020, 132 (48) , 21832-21839. https://doi.org/10.1002/ange.202008413
  23. Xue Gong, Ruomeng Li, Jing Wang, Jie Wei, Kang Ma, Xiaoqing Liu, Fuan Wang. A Smart Theranostic Nanocapsule for Spatiotemporally Programmable Photo‐Gene Therapy. Angewandte Chemie International Edition 2020, 59 (48) , 21648-21655. https://doi.org/10.1002/anie.202008413
  24. Xuchu Wang, Zhihua Tao. Expanding the analytical applications of nucleic acid hybridization using junction probes. Analytical Methods 2020, 12 (41) , 4931-4938. https://doi.org/10.1039/D0AY01605E
  25. Lingjie Ren, Xiaoxia Chen, Chang Feng, Lei Ding, Xiaomin Liu, Tianshu Chen, Fan Zhang, Yanli Li, Zhongliang Ma, Bo Tian, Xiaoli Zhu. Visualized and cascade-enhanced gene silencing by smart DNAzyme-graphene nanocomplex. Nano Research 2020, 13 (8) , 2165-2174. https://doi.org/10.1007/s12274-020-2826-5
  26. Jing Li, Shijun Cai, Bing Zhou, Xiangxian Meng, Qiuping Guo, Xiaohai Yang, Jin Huang, Kemin Wang. Photocaged FRET nanoflares for intracellular microRNA imaging. Chemical Communications 2020, 56 (45) , 6126-6129. https://doi.org/10.1039/D0CC02395G
  27. 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
  28. Zhen Zou, Ziling Luo, Xuan Xu, Sheng Yang, Zhihe Qing, Juewen Liu, Ronghua Yang. Photoactivatable fluorescent probes for spatiotemporal-controlled biosensing and imaging. TrAC Trends in Analytical Chemistry 2020, 125 , 115811. https://doi.org/10.1016/j.trac.2020.115811
  29. Aleksandr A. Spelkov, Ekaterina A. Goncharova, Artemii M. Savin, Dmitry M. Kolpashchikov. Bifunctional RNA‐Targeting Deoxyribozyme Nanodevice as a Potential Theranostic Agent. Chemistry – A European Journal 2020, 26 (16) , 3489-3493. https://doi.org/10.1002/chem.201905528
  30. Maria Hepel. Magnetic Nanoparticles for Nanomedicine. Magnetochemistry 2020, 6 (1) , 3. https://doi.org/10.3390/magnetochemistry6010003
  31. Anzhi Sheng, Lihong Su, Mohammed Jalalah, M. S. Al-Assiri, Farid A. Harraz, Juan Zhang. Hydrazone chemistry assisted DNAzyme for the analysis of double targets. Chemical Communications 2020, 56 (5) , 695-698. https://doi.org/10.1039/C9CC09389C
  32. Orakan Hanpanich, Tomoya Oyanagi, Naohiko Shimada, Atsushi Maruyama. Cationic copolymer-chaperoned DNAzyme sensor for microRNA detection. Biomaterials 2019, 225 , 119535. https://doi.org/10.1016/j.biomaterials.2019.119535
  33. Krishnan, Goud. Magnetic Particle Bioconjugates: A Versatile Sensor Approach. Magnetochemistry 2019, 5 (4) , 64. https://doi.org/10.3390/magnetochemistry5040064
  34. Fumiaki Tomoike, Hiroshi Abe. RNA imaging by chemical probes. Advanced Drug Delivery Reviews 2019, 147 , 44-58. https://doi.org/10.1016/j.addr.2019.08.001
  35. Saira Bakshi, Andrey Zakharchenko, Sergiy Minko, Dmitry Kolpashchikov, Evgeny Katz. Towards Nanomaterials for Cancer Theranostics: A System of DNA-Modified Magnetic Nanoparticles for Detection and Suppression of RNA Marker in Cancer Cells. Magnetochemistry 2019, 5 (2) , 24. https://doi.org/10.3390/magnetochemistry5020024
  36. Zejun Wang, Wenhan Liu, Chunhai Fan, Nan Chen. Visualizing mRNA in live mammalian cells. Methods 2019, 161 , 16-23. https://doi.org/10.1016/j.ymeth.2019.03.008
  37. Yaroslav Filipov, Andrey Zakharchenko, Sergiy Minko, Evgeny Katz. Magneto‐Controlled Biocatalytic Cascades with Logically Processed Input Signals – Substrate Channeling versus Free Diffusion. ChemPhysChem 2018, 19 (22) , 3035-3043. https://doi.org/10.1002/cphc.201800851
  38. Roya Tavallaie, Joshua McCarroll, Marion Le Grand, Nicholas Ariotti, Wolfgang Schuhmann, Eric Bakker, Richard David Tilley, David Brynn Hibbert, Maria Kavallaris, John Justin Gooding. Nucleic acid hybridization on an electrically reconfigurable network of gold-coated magnetic nanoparticles enables microRNA detection in blood. Nature Nanotechnology 2018, 13 (11) , 1066-1071. https://doi.org/10.1038/s41565-018-0232-x
  39. Beibei Shan, Yuhan Pu, Yingfan Chen, Mengling Liao, Ming Li. Novel SERS labels: Rational design, functional integration and biomedical applications. Coordination Chemistry Reviews 2018, 371 , 11-37. https://doi.org/10.1016/j.ccr.2018.05.007
  40. Baggio A. Evangelista, Yoon‐Seong Kim, Dmitry M. Kolpashchikov. FaptaSyme: A Strategy for Converting a Monomer/Oligomer‐Nonselective Aptameric Sensor into an Oligomer‐Selective One. ChemBioChem 2018, 19 (11) , 1123-1126. https://doi.org/10.1002/cbic.201800017
  41. M. Zouari, S. Campuzano, J.M. Pingarrón, N. Raouafi. Ultrasensitive determination of microribonucleic acids in cancer cells with nanostructured-disposable electrodes using the viral protein p19 for recognition of ribonucleic acid/microribonucleic acid homoduplexes. Electrochimica Acta 2018, 262 , 39-47. https://doi.org/10.1016/j.electacta.2017.12.190
  42. Saira F. Bakshi, Nataliia Guz, Andrey Zakharchenko, Han Deng, Alexei V. Tumanov, Craig D. Woodworth, Sergiy Minko, Dmitry M. Kolpashchikov, Evgeny Katz. Nanoreactors based on DNAzyme-functionalized magnetic nanoparticles activated by magnetic field. Nanoscale 2018, 10 (3) , 1356-1365. https://doi.org/10.1039/C7NR08581H
  43. Vaisakh V. Mohanan, Balaram Pradhan, Vimala Sridurai, Channabasaveshwar V. Yelamaggad, Ammathnadu S. Achalkumar, Geetha G. Nair. Giant enhancement and facile tuning of photoluminescence in a soft anisotropic magneto-gel. Nanoscale 2018, 10 (33) , 15686-15695. https://doi.org/10.1039/C8NR02725K
Download PDF

  • Abstract

    High Resolution Image
    Download MS PowerPoint Slide

    Scheme 1

    Scheme 1. Principle of Magnetic Field-Activated Deoxyribozyme Sensora
    High Resolution Image
    Download MS PowerPoint Slide

    Scheme a(A) Binary deoxyribozyme sensor (BiDZ) as reported earlier. (18) DNA strands DZa and DZb hybridize to adjacent position of analyte and form deoxyribozyme catalytic core, which cleaves fluorogenic F-sub and increases sample fluorescence. (B) MaBiDZ developed in this study. Magnetic bead (MaB1)-bound DZa forms a catalytic core with DZb in the presence of analyte. The activated sensor produces signal only when: (i) second species of magnetic beads, MaB2 carrying F-sub is present and (ii) magnetic field that aggregates MaB1 and MaB2 is applied. See the DNA sequences in Table 1.

    Figure 1

    Figure 1. Comparison of in vitro fluorescent response of BiDZ and MaBiDZ sensor systems. (A) Time dependent response of BiDZ and MaBiDZ sensor: (a) without F-sub, (b) without synthetic Twist (see Table1) analyte, (c) response of MaBiDz without magnet applied, (d) response of BiDZ, and (e) MaBiDZ activated with magnetic field in the presence of 1 nM synthetic Twist analyte (see SI for concentrations of all other components of the BiDZ and MaBiDZ probe.) (B) Response of MaBiDZ (d, e, f) compared to BiDZ (a, b, c) in the presence of different concentrations of Twist analyte after 30 (a,d), 60 (b,e) or 120 (c,f) min. All error bars are the result of three independent measurements; some bars are not visible because they are smaller than the labels for the experimental points.

    High Resolution Image
    Download MS PowerPoint Slide

    Figure 2

    Figure 2. Intracellular testing of MaBiDZ sensor. CLSM images of (A) Twist-overexpressing MCF-7 cancer cells treated with MaBiDZ sensor with magnetic field applied and (C) no magnetic field applied. Analogously treated cervical epithelial cells (expressing low levels of Twist) with (B) magnetic field applied and (D) without magnetic field. Images were taken after 2.5 h of incubation time. Nuclei are stained with Hoechst nuclear stain and visualized with 408 nm laser. Surfaces are stained with anti-epithelial cell adhesion molecule (EpCAM) antibody and visualized with a 635 nm laser. Fluorescence from the MaBiDZ probe is visualized with the 488 nm laser. Corresponding flow cytometry data are shown as insets below each image. The gates on flow cytometry plots indicate percent of EpCAM positive cells with low and high MaBiDZ fluorescence. The number of internalized particles was estimated to be ca. 1 × 106 MaBiDZ per cell (see Figure SI11). Scale bar is 20 μm.

    High Resolution Image
    Download MS PowerPoint Slide

  • References

    ARTICLE SECTIONS
    Jump To

    This article references 25 other publications.

    1. 1
      Germond, A.; Fujita, H.; Ichimura, T.; Watanabe, T. M. Biophys. Rev. 2016, 8, 121– 138 DOI: 10.1007/s12551-016-0195-9
      Google Scholar
      1
      Design and development of genetically encoded fluorescent sensors to monitor intracellular chemical and physical parameters
      Germond, Arno; Fujita, Hideaki; Ichimura, Taro; Watanabe, Tomonobu M.
      Biophysical Reviews (2016), 8 (2), 121-138CODEN: BRIECG; ISSN:1867-2450. (Springer)
      Over the past decades many researchers have made major contributions towards the development of genetically encoded (GE) fluorescent sensors derived from fluorescent proteins. GE sensors are now used to study biol. phenomena by facilitating the measurement of biochem. behaviors at various scales, ranging from single mols. to single cells or even whole animals. Here, we review the historical development of GE fluorescent sensors and report on their current status. We specifically focus on the development strategies of the GE sensors used for measuring pH, ion concns. (e.g., chloride and calcium), redox indicators, membrane potential, temp., pressure, and mol. crowding. We demonstrate that these fluroescent protein-based sensors have a shared history of concepts and development strategies, and we highlight the most original concepts used to date. We believe that the understanding and application of these various concepts will pave the road for the development of future GE sensors and lead to new breakthroughs in bioimaging.
      >> More from SciFinder ®
      https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC28XnsFWntrg%253D&md5=046fcb72eeb3bd731b0691767c475d8e
    2. 2
      (a) Cui, M.; You, L.; Ren, X.; Zhao, W.; Liao, Q.; Zhao, Y. Biochem. Biophys. Res. Commun. 2016, 471, 10– 14 DOI: 10.1016/j.bbrc.2015.12.101
      Google Scholar
      There is no corresponding record for this reference.
      (b) Guo, D.; Barry, L.; Lin, S. S. H.; Huang, V.; Li, L.-C. RNA Biol. 2014, 11, 1221– 1225 DOI: 10.4161/15476286.2014.972853
      Google Scholar
      2b
      RNAa in action: from the exception to the norm
      Guo Dan; Barry Liam; Lin Sharon Szu Hua; Huang Vera; Li Long-Cheng
      RNA biology (2014), 11 (10), 1221-5 ISSN:.
      Small RNA programmed Argonautes are sophisticated cellular effector platforms known to be involved in a diverse array of functions ranging from mRNA cleavage, translational inhibition, DNA elimination, epigenetic silencing, alternative splicing and even gene activation. First observed in human cells, small RNA-induced gene activation, also known as RNAa, involves the targeted recruitment of Argonaute proteins to specific promoter sequences followed by induction of stable epigenetic changes which promote transcription. The existence of RNAa remains contentious due to its elusive mechanism. A string of recent studies in C. elegans provides unequivocal evidence for RNAa's fundamental role in sculpting the epigenetic landscape and maintaining active transcription of endogenous genes and supports the presence of a functionally sophisticated network of small RNA-Argonaute pathways consisting of opposite yet complementary "yin and yang" regulatory elements. In this review, we summarize key findings from recent studies of endogenous RNAa in C. elegans, with an emphasis on the Argonaute protein CSR-1.
      >> More from SciFinder ®
      https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A280%3ADC%252BC2MvnslWnsg%253D%253D&md5=0fce92a5a3a495558ec8458e418482af
      (c) Qi, X.; Zhang, D.-H.; Wu, N.; Xiao, J.-H.; Wang, X.; Ma, W. J. Med. Genet. 2015, 52, 710– 718 DOI: 10.1136/jmedgenet-2015-103334
      Google Scholar
      2c
      ceRNA in cancer: possible functions and clinical implications
      Qi Xiaolong; Zhang Da-Hong; Wu Nan; Xiao Jun-Hua; Wang Xiang; Ma Wang
      Journal of medical genetics (2015), 52 (10), 710-8 ISSN:.
      Competing endogenous RNAs (ceRNAs) are transcripts that can regulate each other at post-transcription level by competing for shared miRNAs. CeRNA networks link the function of protein-coding mRNAs with that of non-coding RNAs such as microRNA, long non-coding RNA, pseudogenic RNA and circular RNA. Given that any transcripts harbouring miRNA response element can theoretically function as ceRNAs, they may represent a widespread form of post-transcriptional regulation of gene expression in both physiology and pathology. CeRNA activity is influenced by multiple factors such as the abundance and subcellular localisation of ceRNA components, binding affinity of miRNAs to their sponges, RNA editing, RNA secondary structures and RNA-binding proteins. Aberrations in these factors may deregulate ceRNA networks and thus lead to human diseases including cancer. In this review, we introduce the mechanisms and molecular bases of ceRNA networks, discuss their roles in the pathogenesis of cancer as well as methods of predicting and validating ceRNA interplay. At last, we discuss the limitations of current ceRNA theory, propose possible directions and envision the possibilities of ceRNAs as diagnostic biomarkers or therapeutic targets.
      >> More from SciFinder ®
      https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A280%3ADC%252BC283htV2htQ%253D%253D&md5=0489dc0a4a01909899f8c39a83130e9d
    3. 3
      Bertrand, E.; Chartrand, P.; Schaefer, M.; Shenoy, S. M.; Singer, R. H.; Long, R. M. Mol. Cell 1998, 2, 437– 445 DOI: 10.1016/S1097-2765(00)80143-4
      Google Scholar
      There is no corresponding record for this reference.
    4. 4
      Paige, J. S.; Wu, K. Y.; Jaffrey, S. R. Science 2011, 333, 642– 646 DOI: 10.1126/science.1207339
      Google Scholar
      4
      RNA mimics of green fluorescent protein
      Paige, Jeremy S.; Wu, Karen Y.; Jaffrey, Samie R.
      Science (Washington, DC, United States) (2011), 333 (6042), 642-646CODEN: SCIEAS; ISSN:0036-8075. (American Association for the Advancement of Science)
      Green fluorescent protein (GFP) and its derivs. have transformed the use and anal. of proteins for diverse applications. Like proteins, RNA has complex roles in cellular function and is increasingly used for various applications, but a comparable approach for fluorescently tagging RNA is lacking. Here, we describe the generation of RNA aptamers that bind fluorophores resembling the fluorophore in GFP. These RNA-fluorophore complexes create a palette that spans the visible spectrum. An RNA-fluorophore complex, termed Spinach, resembles enhanced GFP and emits a green fluorescence comparable in brightness with fluorescent proteins. Spinach is markedly resistant to photobleaching, and Spinach fusion RNAs can be imaged in living cells. These RNA mimics of GFP provide an approach for genetic encoding of fluorescent RNAs.
      >> More from SciFinder ®
      https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3MXpt1yisbw%253D&md5=dc757f578e744c78062a68b52d15a961
    5. 5
      (a) Holtzer, L.; Oleinich, I.; Anzola, M.; Lindberg, E.; Sadhu, K. K.; Gonzalez-Gaitan, M.; Winssinger, N. ACS Cent. Sci. 2016, 2, 394– 400 DOI: 10.1021/acscentsci.6b00054
      Google Scholar
      5a
      Nucleic Acid Templated Chemical Reaction in a Live Vertebrate
      Holtzer, Laurent; Oleinich, Igor; Anzola, Marcello; Lindberg, Eric; Sadhu, Kalyan K.; Gonzalez-Gaitan, Marcos; Winssinger, Nicolas
      ACS Central Science (2016), 2 (6), 394-400CODEN: ACSCII; ISSN:2374-7951. (American Chemical Society)
      Nucleic acid templated reactions are enabled by the hybridization of probe-reagent conjugates resulting in high effective reagent concn. and fast chem. transformation. We have developed a reaction that harnesses cellular microRNA (miRNA) to yield the cleavage of a linker releasing fluorogenic rhodamine in a live vertebrate. The reaction is based on the catalytic photoredn. of an azide by a ruthenium complex. We showed that this system reports specific expression of miRNA in living tissues of a vertebrate.
      >> More from SciFinder ®
      https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC28XotFelsLo%253D&md5=241e3e5dcd20cdf43600dd27137b9111
      (b) Michaelis, J.; Roloff, A.; Seitz, O. Org. Biomol. Chem. 2014, 12, 2821– 2833 DOI: 10.1039/C4OB00096J
      Google Scholar
      There is no corresponding record for this reference.
      (c) Franzini, R. M.; Kool, E. T. Bioconjugate Chem. 2011, 22, 1869– 1877 DOI: 10.1021/bc2003567
      Google Scholar
      5c
      Improved Templated Fluorogenic Probes Enhance the Analysis of Closely Related Pathogenic Bacteria by Microscopy and Flow Cytometry
      Franzini, Raphael M.; Kool, Eric T.
      Bioconjugate Chemistry (2011), 22 (9), 1869-1877CODEN: BCCHES; ISSN:1043-1802. (American Chemical Society)
      Templated fluorescence activation has recently emerged as a promising mol. approach to detect and differentiate nucleic acid sequences in vitro and in cells. Here, we describe the application of a reductive quencher release strategy to the taxonomic anal. of Gram-neg. bacteria by targeting a single nucleotide difference in their 16S rRNA in a two-color assay. For this purpose, it was necessary to develop a release linker contg. a quencher suitable for red and near-IR fluorophores, and to improve methods for the delivery of probes into cells. A cyanine-dye labeled oligonucleotide probe contg. the new quencher-release linker showed unprecedentedly low background signal and high fluorescence turn-on ratios. The combination of a fluorescein-contg. and a near-IR emitting probe discriminated E. coli from S. enterica despite nearly identical ribosomal target sequences. Two-color anal. by microscopy and the first successful discrimination of bacteria by two-color flow cytometry with templated reactive probes are described.
      >> More from SciFinder ®
      https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3MXhtVOktrbK&md5=92ba463514226e5eda1ed0cdf39be94f
    6. 6
      Santangelo, P. J.; Nix, B.; Tsourkas, A.; Bao, G. Nucleic Acids Res. 2004, 32, e57 DOI: 10.1093/nar/gnh062
      Google Scholar
      6
      Dual FRET molecular beacons for mRNA detection in living cells
      Santangelo, Philip J.; Nix, Brent; Tsourkas, Andrew; Bao, Gang
      Nucleic Acids Research (2004), 32 (6), e57/1-e57/9CODEN: NARHAD; ISSN:0305-1048. (Oxford University Press)
      The ability to visualize in real-time the expression level and localization of specific endogenous RNAs in living cells can offer tremendous opportunities for biol. and disease studies. Here we demonstrate such a capability using a pair of mol. beacons, one with a donor and the other with an acceptor fluorophore that hybridize to adjacent regions on the same mRNA target, resulting in fluorescence resonance energy transfer (FRET). Detection of the FRET signal significantly reduced false positives, leading to sensitive imaging of K-ras and survivin mRNAs in live HDF and MIAPaCa-2 cells. FRET detection gave a ratio of 2.25 of K-ras mRNA expression in stimulated and unstimulated HDF, comparable to the ratio of 1.95 using RT-PCR, and in contrast to the single-beacon result of 1.2. We further revealed intriguing details of K-ras and survivin mRNA localization in living cells. The dual FRET mol. beacons approach provides a novel technique for sensitive RNA detection and quantification in living cells.
      >> More from SciFinder ®
      https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD2cXjt1ejsb8%253D&md5=3a808296da2f8bc0ad8f5b6bc20e5f7f
    7. 7
      (a) Zhao, D.; Yang, Y.; Qu, N.; Chen, M.; Ma, Z.; Krueger, C. J.; Behlke, M. A.; Chen, A. K. Biomaterials 2016, 100, 172– 183 DOI: 10.1016/j.biomaterials.2016.05.022
      Google Scholar
      7a
      Single-molecule detection and tracking of RNA transcripts in living cells using phosphorothioate-optimized 2'-O-methyl RNA molecular beacons
      Zhao, Dan; Yang, Yantao; Qu, Na; Chen, Mingming; Ma, Zhao; Krueger, Christopher J.; Behlke, Mark A.; Chen, Antony K.
      Biomaterials (2016), 100 (), 172-183CODEN: BIMADU; ISSN:0142-9612. (Elsevier Ltd.)
      Mol. beacons (MBs) composed of 2'-O-Me RNA (2Me) and phosphorothioate (PS) linkages throughout the backbone (2Me/PSFULL MBs) have enabled long-term imaging of RNA in living cells, but excess PS modification can induce nonspecific binding, causing false-pos. signals. In this study, we evaluate the intracellular stability of MBs composed of 2Me with various PS modifications, and found that false-pos. signals could be reduced to marginal levels when the MBs possess a fully PS-modified loop domain and a phosphodiester stem (2Me/PSLOOP MB). Addnl., 2Me/PSLOOP MBs exhibited uncompromised hybridization kinetics, prolonged functionality and >88% detection accuracy for single RNA transcripts, and could do so without interfering with gene expression or cell growth. Finally, 2Me/PSLOOP MBs could image the dynamics of single mRNA transcripts in the nucleus and the cytoplasm simultaneously, regardless of whether the MBs targeted the 5'- or the 3'-UTR. Together, these findings demonstrate the effectiveness of loop-domain PS modification in reducing nonspecific signals and the potential for sensitive and accurate imaging of individual RNAs at the single-mol. level. With the growing interest in the role of RNA localization and dynamics in health and disease, 2Me/PSLOOP MBs could enable new discoveries in RNA research.
      >> More from SciFinder ®
      https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC28Xos12rtbk%253D&md5=1c70b0aa6afd761532be7911f168601d
      (b) Giraldo-Vela, J. P.; Kang, W.; McNaughton, R. L.; Zhang, X.; Wile, B. M.; Tsourkas, A.; Bao, G.; Espinosa, H. D. Small 2015, 11, 2386– 2391 DOI: 10.1002/smll.201401137
      Google Scholar
      There is no corresponding record for this reference.
    8. 8
      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
      8
      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.
      >> More from SciFinder ®
      https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD2sXhtlKru7jE&md5=71e766b235655781c09ccfc331b37762
      (a) Prigodich, A. E.; Seferos, D. S.; Massich, M. D.; Giljohann, D. A.; Lane, B. C.; Mirkin, C. A. ACS Nano 2009, 3, 2147– 2152 DOI: 10.1021/nn9003814
      Google Scholar
      There is no corresponding record for this reference.
      (b) Zheng, D.; Seferos, D. S.; Giljohann, D. A.; Patel, P. C.; Mirkin, C. A. Nano Lett. 2009, 9, 3258– 3261 DOI: 10.1021/nl901517b
      Google Scholar
      8b
      Aptamer Nano-flares for Molecular Detection in Living Cells
      Zheng, Dan; Seferos, Dwight S.; Giljohann, David A.; Patel, Pinal C.; Mirkin, Chad A.
      Nano Letters (2009), 9 (9), 3258-3261CODEN: NALEFD; ISSN:1530-6984. (American Chemical Society)
      The authors demonstrate a composite nanomaterial, termed an aptamer nano-flare, that can directly quantify an intracellular analyte in a living cell. Aptamer nano-flares consist of a gold nanoparticle core functionalized with a dense monolayer of nucleic acid aptamers with a high affinity for ATP. The probes bind selectively to target mols. and release fluorescent reporters which indicate the presence of the analyte. Addnl., these nanoconjugates are readily taken up by cells where their signal intensity can be used to quantify intracellular analyte concn. These nanoconjugates are a promising approach for the intracellular quantification of other small mols. or proteins, or as agents that use aptamer binding to elicit a biol. response in living systems.
      >> More from SciFinder ®
      https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD1MXptlSmtbw%253D&md5=ed0a131b9ccca7002b24570f3032628b
      (c) Pan, W.; Li, Y.; Wang, M.; Yang, H.; Li, N.; Tang, B. Chem. Commun. 2016, 52, 4569– 4572 DOI: 10.1039/C5CC10147F
      Google Scholar
      There is no corresponding record for this reference.
    9. 9
      Briley, W. E.; Bondy, M. H.; Randeria, P. S.; Dupper, T. J.; Mirkin, C. A. Proc. Natl. Acad. Sci. U. S. A. 2015, 112, 9591– 9595 DOI: 10.1073/pnas.1510581112
      Google Scholar
      There is no corresponding record for this reference.
    10. 10
      (a) Feyder, M.; Goff, L. A. J. Clin. Invest. 2016, 126, 2783– 2791 DOI: 10.1172/JCI84422
      Google Scholar
      There is no corresponding record for this reference.
      (b) Ouellet, J. Front. Chem. 2016, 429 DOI: 10.3389/fchem.2016.00029
      Google Scholar
      10b
      RNA Fluorescence with Light-Up Aptamers
      Ouellet Jonathan
      Frontiers in chemistry (2016), 4 (), 29 ISSN:2296-2646.
      Seeing is not only believing; it also includes understanding. Cellular imaging with GFP in live cells has been transformative in many research fields. Modulation of cellular regulation is tightly regulated and innovative imaging technologies contribute to further understand cellular signaling and physiology. New types of genetically encoded biosensors have been developed over the last decade. They are RNA aptamers that bind with their cognate fluorogen ligands and activate their fluorescence. The emergence and the evolution of these RNA aptamers as well as their conversion into a wide spectrum of applications are examined in a global way.
      >> More from SciFinder ®
      https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A280%3ADC%252BC2s3jsFanug%253D%253D&md5=ebf0cfee8a0de42cbef6c299822eb033
      (c) Shigeto, H.; Nakatsuka, K.; Ikeda, T.; Hirota, R.; Kuroda, A.; Funabashi, H. Anal. Chem. 2016, 88, 7894– 7898 DOI: 10.1021/acs.analchem.6b02710
      Google Scholar
      There is no corresponding record for this reference.
      (d) Takahashi, K.; Ito, S.; Nakamoto, K.; Ito, Y.; Ueno, Y. J. Org. Chem. 2015, 80, 8561– 8570 DOI: 10.1021/acs.joc.5b01132
      Google Scholar
      There is no corresponding record for this reference.
      (e) Urbanek, M. O.; Galka-Marciniak, P.; Olejniczak, M.; Krzyzosiak, W. J. RNA Biol. 2014, 11, 1083– 1095 DOI: 10.4161/rna.35506
      Google Scholar
      10e
      RNA imaging in living cells - methods and applications
      Urbanek Martyna O; Galka-Marciniak Paulina; Olejniczak Marta; Krzyzosiak Wlodzimierz J
      RNA biology (2014), 11 (8), 1083-95 ISSN:.
      Numerous types of transcripts perform multiple functions in cells, and these functions are mainly facilitated by the interactions of the RNA with various proteins and other RNAs. Insight into the dynamics of RNA biosynthesis, processing and cellular activities is highly desirable because this knowledge will deepen our understanding of cell physiology and help explain the mechanisms of RNA-mediated pathologies. In this review, we discuss the live RNA imaging systems that have been developed to date. We highlight information on the design of these systems, briefly discuss their advantages and limitations and provide examples of their numerous applications in various organisms and cell types. We present a detailed examination of one application of RNA imaging systems: this application aims to explain the role of mutant transcripts in human disease pathogenesis caused by triplet repeat expansions. Thus, this review introduces live RNA imaging systems and provides a glimpse into their various applications.
      >> More from SciFinder ®
      https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A280%3ADC%252BC2MzmtVSgsg%253D%253D&md5=00375391046f0df6d39ca4dc084e0eeb
    11. 11
      (a) Wang, C.; Zhu, Z.; Song, Y.; Lin, H.; Yang, C. J.; Tan, W. Chem. Commun. 2011, 47, 5708– 5710 DOI: 10.1039/c1cc10481k
      Google Scholar
      There is no corresponding record for this reference.
      (b) Joshi, K. B.; Vlachos, A.; Mikat, V.; Deller, T.; Heckel, A. Chem. Commun. 2012, 48, 2746– 2748 DOI: 10.1039/C2CC16654B
      Google Scholar
      There is no corresponding record for this reference.
      (c) Rinne, J. S.; Kaminski, T. P.; Kubitscheck, U.; Heckel, A. Chem. Commun. 2013, 49, 5375– 5377 DOI: 10.1039/c3cc42420k
      Google Scholar
      There is no corresponding record for this reference.
    12. 12
      (a) Ruble, B. K.; Yeldell, S. B.; Dmochowski, I. J. J. Inorg. Biochem. 2015, 150, 182– 188 DOI: 10.1016/j.jinorgbio.2015.03.010
      Google Scholar
      There is no corresponding record for this reference.
      (b) Tang, X.; Zhang, J.; Sun, J.; Wang, Y.; Wu, J.; Zhang, L. Org. Biomol. Chem. 2013, 11, 7814– 7824 DOI: 10.1039/c3ob41735b
      Google Scholar
      There is no corresponding record for this reference.
      (c) Bort, G.; Gallavardin, T.; Ogden, D.; Dalko, P. I. Angew. Chem., Int. Ed. 2013, 52, 4526– 4537 DOI: 10.1002/anie.201204203
      Google Scholar
      12c
      From One-Photon to Two-Photon Probes: "Caged" Compounds, Actuators, and Photoswitches
      Bort, Guillaume; Gallavardin, Thibault; Ogden, David; Dalko, Peter I.
      Angewandte Chemie, International Edition (2013), 52 (17), 4526-4537CODEN: ACIEF5; ISSN:1433-7851. (Wiley-VCH Verlag GmbH & Co. KGaA)
      A review. Mol. systems that can be remotely controlled by light are gaining increasing importance in cell biol., physiol., and neurosciences because of the spatial and temporal precision that is achievable with laser microscopy. Two-photon excitation has significant advantages deep in biol. tissues, but raises problems in the design of "smart" probes compatible with cell physiol. This Review discusses the chem. challenges in generating suitable two-photon probes.
      >> More from SciFinder ®
      https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3sXislWgsb4%253D&md5=0d8affb2b63341dcbad735299c2464ec
    13. 13
      (a) Luo, D.; Saltzman, W. M. Nat. Biotechnol. 2000, 18, 33– 37 DOI: 10.1038/71889
      Google Scholar
      There is no corresponding record for this reference.
      (b) Patil, S. D.; Rhodes, D. G.; Burgess, D. J. AAPS J. 2005, 7, E61– 77 DOI: 10.1208/aapsj070109
      Google Scholar
      There is no corresponding record for this reference.
      (c) Bishop, C. J.; Kozielski, K. L.; Green, J. J. J. Controlled Release 2015, 219, 488– 499 DOI: 10.1016/j.jconrel.2015.09.046
      Google Scholar
      There is no corresponding record for this reference.
    14. 14
      Dobson, J. Nat. Nanotechnol. 2008, 3, 139– 143 DOI: 10.1038/nnano.2008.39
      Google Scholar
      14
      Remote control of cellular behaviour with magnetic nanoparticles
      Dobson, Jon
      Nature Nanotechnology (2008), 3 (3), 139-143CODEN: NNAABX; ISSN:1748-3387. (Nature Publishing Group)
      A review. By binding magnetic nanoparticles to the surface of cells, it is possible to manipulate and control cell function with an external magnetic field. The technique of activating cells with magnetic nanoparticles offers a means to isolate and explore cellular mechanics and ion channel activation to gain better understanding of these processes. Here, we go beyond using this technique as an investigative tool and focus on its potential to actively control cellular functions and processes with an eye towards biol. and clin. applications. In particular, we focus on applications in tissue engineering and regenerative medicine.
      >> More from SciFinder ®
      https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD1cXivFKgtrY%253D&md5=700c579988781da1d25c69e9023eb705
    15. 15
      (a) Wang, N.; Butler, J. P.; Ingber, D. E. Science 1993, 260, 1124– 1127 DOI: 10.1126/science.7684161
      Google Scholar
      There is no corresponding record for this reference.
      (b) Meyer, C. J.; Alenghat, F. J.; Rim, P.; Fong, J. H.-J.; Fabry, B.; Ingber, D. E. Nat. Cell Biol. 2000, 2, 666– 668 DOI: 10.1038/35023621
      Google Scholar
      15b
      Mechanical control of cyclic AMP signalling and gene transcription through integrins
      Meyer, Christian J.; Alenghat, Francis J.; Rim, Paul; Fong, Jenny Hwai-Jen; Fabry, Ben; Ingber, Donald E.
      Nature Cell Biology (2000), 2 (9), 666-668CODEN: NCBIFN; ISSN:1465-7392. (Nature Publishing Group)
      This study was carried out to discriminate between two alternative hypotheses as to how cells sense mech. forces and transduce them into changes in gene transcription. Do cells sense mech. signals through generalized membrane distortion or through specific transmembrane receptors, such as integrins. Here the authors show that mech. stresses applied to the cell surface alter the cAMP signalling cascade and downstream gene transcription by modulating local release of signals generated by activated integrin receptors in a G-protein-dependent manner, whereas distortion of integrins in the absence of receptor occupancy has no effect.
      >> More from SciFinder ®
      https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD3cXmslGks7k%253D&md5=033770918a88216173175fb8aaec4e52
    16. 16
      (a) Bausch, A. R.; Hellerer, U.; Essler, M.; Aepfelbacher, M.; Sackmann, E. Biophys. J. 2001, 80, 2649– 2657 DOI: 10.1016/S0006-3495(01)76234-0
      Google Scholar
      There is no corresponding record for this reference.
      (b) Bausch, A. R.; Möller, W.; Sackmann, E. Biophys. J. 1999, 76, 573– 579 DOI: 10.1016/S0006-3495(99)77225-5
      Google Scholar
      16b
      Measurement of local viscoelasticity and forces in living cells by magnetic tweezers
      Bausch, Andreas R.; Moller, Winfried; Sackmann, Erich
      Biophysical Journal (1999), 76 (1, Pt. 1), 573-579CODEN: BIOJAU; ISSN:0006-3495. (Biophysical Society)
      We measured the viscoelastic properties of the cytoplasm of J774 macrophages with a recently developed microrheometer. Ferromagnetic beads (1.3 μm in diam.) were used to det. the local viscoelastic moduli. Step-force pulses were applied to the magnetic beads and the displacement was obsd. by single particle tracking. By analyzing the creep response curves in terms of a triphasic mech. equiv. circuit, we measured the shear elastic modulus, the effective viscosities, and the strain relaxation time. The values of the shear modulus vary by more than an order of magnitude within the cell population (range, 20-735 Pa; av., 343 Pa) and by a factor of 2 within single cells. The effective viscosity of the cytoplasm exhibits a relatively sharp distribution about an av. of η = 210 Pa s (± 143 Pa s). We measured the displacement field generated by the local forces by observing the induced motion of nonmagnetic beads. Even at distances of the order of 1 μm, no induced motion was seen, suggesting that the cytoplasm is composed of clusters of densely packed and cross-linked filaments sepd. by soft regions. In another series of expts. we analyzed the magnetophoretic motion of the ferromagnetic beads at a const. magnetic force. Measuring the bead velocity parallel and perpendicular to the applied force showed that local active forces on the beads varied from 50 to 900 pN.
      >> More from SciFinder ®
      https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADyaK1MXjs1Sjug%253D%253D&md5=c7cfba90639225899ff9ac732ecdc983
      (c) Glogauer, M.; Ferrier, J. Pfluegers Arch. 1997, 435, 320– 327 DOI: 10.1007/s004240050518
      Google Scholar
      There is no corresponding record for this reference.
    17. 17
      Hughes, S.; McBain, S.; Dobson, J.; El Haj, A. J. J. J. R. Soc., Interface 2008, 5, 855– 863 DOI: 10.1098/rsif.2007.1274
      Google Scholar
      17
      Selective activation of mechanosensitive ion channels using magnetic particles
      Hughes, Steven; McBain, Stuart; Dobson, Jon; El Haj, Alicia J.
      Journal of the Royal Society, Interface (2008), 5 (25), 855-863CODEN: JRSICU; ISSN:1742-5689. (Royal Society)
      This study reports the preliminary development of a novel magnetic particle-based technique that permits the application of highly localized mech. forces directly to specific regions of an ion-channel structure. We demonstrate that this approach can be used to directly and selectively activate a mechanosensitive ion channel of interest, namely TREK-1. It is shown that manipulation of particles targeted against the extended extracellular loop region of TREK-1 leads to changes in whole-cell currents consistent with changes in TREK-1 activity. Responses were absent when particles were coated with RGD (Arg-Gly-Asp) peptide or when magnetic fields were applied in the absence of magnetic particles. It is concluded that changes in whole-cell current are the result of direct force application to the extracellular loop mechano-gating. It is hypothesized that the extended loop region of TREK-1 may act as a tension spring that acts to regulate sensitivity to mech. forces, in a nature similar to that described for MscL. The development of a technique that permits the direct manipulation of mechanosensitive ion channels in real time without the need for pharmacol. drugs has huge potential benefits not only for basic biol. research of ion-channel gating mechanisms, but also potentially as a tool for the treatment of human diseases caused by ion-channel dysfunction.
      >> More from SciFinder ®
      https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD1cXpvVGntLc%253D&md5=a114c2e9f738827ec18e4e5c25a68f75
    18. 18
      (a) Kolpashchikov, D. M. ChemBioChem 2007, 8, 2039– 2042 DOI: 10.1002/cbic.200700384
      Google Scholar
      There is no corresponding record for this reference.
      (b) Mokany, E.; Bone, S. M.; Young, P. E.; Doan, T. B.; Todd, A. V. J. Am. Chem. Soc. 2010, 132, 1051– 1059 DOI: 10.1021/ja9076777
      Google Scholar
      18b
      MNAzymes, a Versatile New Class of Nucleic Acid Enzymes That Can Function as Biosensors and Molecular Switches
      Mokany, Elisa; Bone, Simon M.; Young, Paul E.; Doan, Tram B.; Todd, Alison V.
      Journal of the American Chemical Society (2010), 132 (3), 1051-1059CODEN: JACSAT; ISSN:0002-7863. (American Chemical Society)
      To increase the versatility and utility of nucleic acid enzymes, multicomponent complexes known as MNAzymes were developed, which produce amplified output signals in response to specific input signals. Multiple oligonucleotide partzymes assemble into active MNAzymes only in the presence of an input assembly facilitator such as a target nucleic acid. Once formed, MNAzymes catalytically modify a generic substrate, generating an amplified output signal that heralds the presence of the target while leaving the target intact. Several applications were demonstrated, including sensitive, isothermal target detection; discrimination of polymorphisms; and highly specific monitoring of real-time polymerase chain reaction (PCR). Furthermore, their capacity to function as mol. switches and to work in series to create a mol. cascade was demonstrated. The modular nature of MNAzymes, together with the sepn. of input and output functionalities, provides potential for their integration into diverse devices such as diagnostic biosensors, mol. computers, and/or nanoscale machines.
      >> More from SciFinder ®
      https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD1MXhs1WjsLvF&md5=80a44a8993aa11039a48e664af73d761
      (c) Gerasimova, Y. V.; Cornett, E.; Kolpashchikov, D. M. ChemBioChem 2010, 11, 811– 817 DOI: 10.1002/cbic.201000006
      Google Scholar
      There is no corresponding record for this reference.
      (d) Gerasimova, Y. V.; Kolpashchikov, D. M. Angew. Chem., Int. Ed. 2013, 52, 10586– 10588 DOI: 10.1002/anie.201303919
      Google Scholar
      There is no corresponding record for this reference.
    19. 19
      Bumb, A.; Brechbiel, M. W.; Choyke, P. L.; Fugger, L.; Eggeman, A.; Prabhakaran, D.; Hutchinson, J.; Dobson, P. J. Nanotechnology 2008, 19335601 DOI: 10.1088/0957-4484/19/33/335601
      Google Scholar
      There is no corresponding record for this reference.
    20. 20
      (a) Gref, R.; Lück, M.; Quellec, P.; Marchand, M.; Dellacherie, E.; Harnisch, S.; Blunk, T.; Müller, R. H. Colloids Surf., B 2000, 18, 301– 313 DOI: 10.1016/S0927-7765(99)00156-3
      Google Scholar
      There is no corresponding record for this reference.
    21. 21
      Cheng, G. Z.; Chan, J.; Wang, Q.; Zhang, W. Z.; Sun, C. D.; Wang, L. H. Cancer Res. 2007, 67, 1979– 1987 DOI: 10.1158/0008-5472.CAN-06-1479
      Google Scholar
      There is no corresponding record for this reference.
    22. 22
      Watanabe, O.; Imamura, H.; Shimizu, T.; Kinoshita, J.; Okabe, T.; Hirano, A.; Yoshimatsu, K.; Konno, S.; Aiba, M.; Ogawa, K. Anticancer Res. 2004, 24, 3851– 3856
      Google Scholar
      22
      Expression of Twist and Wnt in human breast cancer
      Watanabe, Osamu; Imamura, Hiroshi; Shimizu, Tadao; Kinoshita, Jun; Okabe, Toshihiro; Hirano, Akira; Yoshimatsu, Kazuhiko; Konno, Soichi; Aiba, Motohiko; Ogawa, Kenji
      Anticancer Research (2004), 24 (6), 3851-3856CODEN: ANTRD4; ISSN:0250-7005. (International Institute of Anticancer Research)
      Twist, a transcription factor of the basic helix-loop-helix class, has been suggested to have oncogenic properties. We reported Twist expression was regulated by Wnt/β-catenin signaling and that both Wnt-1 and Twist could contribute to mammary tumorigenesis. The aim of this study was to demonstrate the expression of Twist, Wnt-1 and Wnt-2 in human breast cancer tissue. We examd. the expression in patients with breast cancer by RT-PCR and immunohistochem. RT-PCR of twenty-three pairs of cancer and normal breast tissue revealed that Twist was up-regulated in 69.6% (16/23) of the cancer lesions and 21.7% (5/23) of the normal breast tissues. Wnt-2 was up-regulated in all of the cancer lesions and 13.0% (3/23) of the normal breast tissues, whereas Wnt-1 was expressed in both the cancer and normal breast tissues of the five cases examd. Immunohistochem. analyses revealed that Twist was pos. expressed in 52.2% (12/23) of the cancer lesions and 34.8% (8/23) of the normal breast tissues. Twist and Wnt-2 are highly expressed in breast cancer tissue, suggesting that both mols. could play important roles in mammary carcinogenesis.
      >> More from SciFinder ®
      https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD2MXitFylsLk%253D&md5=23684de587fa14fb044471b29d846ab0
    23. 23
      Li, Y.; Wang, W.; Wang, W.; Yang, R.; Wang, T.; Su, T.; Weng, D.; Tao, T.; Li, W.; Ma, D.; Wang, S. Gynecol. Oncol. 2012, 124, 112– 118 DOI: 10.1016/j.ygyno.2011.09.003
      Google Scholar
      23
      Correlation of TWIST2 up-regulation and epithelial-mesenchymal transition during tumorigenesis and progression of cervical carcinoma
      Li, Yan; Wang, Wei; Wang, Wenwen; Yang, Runfeng; Wang, Tian; Su, Tiefen; Weng, Danhui; Tao, Tao; Li, Wei; Ma, Ding; Wang, Shixuan
      Gynecologic Oncology (2012), 124 (1), 112-118CODEN: GYNOA3; ISSN:0090-8258. (Elsevier B.V.)
      Globally, cervical cancer is the second most common cancer among women, and detg. potential targets involved in tumor progression is necessary. This study investigated the clinic-pathol. significance of twist homolog 2 (TWIST2), a basic helix-loop-helix transcription factor, and correlated TWIST2 and E-cadherin expression in cervical cancer. A series of 142 samples, including 14 cases of normal cervical tissues, 58 cases of cervical intraepithelial neoplasia (CIN) and 70 cases of squamous cell carcinoma (SCC), were examd. TWIST2 and E-cadherin immunohistochem. staining and statistical anal. Increased cytoplasmic and nuclear expression levels of TWIST2 were assocd. with the malignant transformation of cervical epithelium and the histol. progression of cervical cancer. A logistic test showed that TWIST2 was a relatively independent predictor of lymph node metastasis of SCC. Further, increased levels of TWIST2 were also assocd. with aberrant expression of E-cadherin, an important EMT indicator. The present data suggest that TWIST2 overexpression was significantly linked to cervical cancer progression, which makes it a promising marker for detg. the metastatic potential of cervical cancer, and up-regulation of TWIST2, in combination with aberrant E-cadherin expression in primary cervical cancer tissues, may predict the malignant transformation and distal metastasis of carcinomas.
      >> More from SciFinder ®
      https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3MXhsF2rtLjL&md5=8258c2465a996782d558c320554942aa
    24. 24
      Plank, C.; Schillinger, U.; Scherer, F.; Bergemann, C.; Rémy, J.-S.; Krötz, F.; Anton, M.; Lausier, J.; Rosenecker, J. Biol. Chem. 2003, 384, 737– 747 DOI: 10.1515/BC.2003.082
      Google Scholar
      There is no corresponding record for this reference.
    25. 25
      Nguyen, J.; Szoka, F. Acc. Chem. Res. 2012, 45, 1153– 1162 DOI: 10.1021/ar3000162
      Google Scholar
      There is no corresponding record for this reference.

  • Supporting Information

    Supporting Information

    ARTICLE SECTIONS
    Jump To

    The Supporting Information is available free of charge on the ACS Publications website at DOI: 10.1021/jacs.7b06022.

    • Details of experimental procedures, complete structure of probe-analyte complex, and control experiments with cell culture (PDF)


    Terms & Conditions

    Most electronic Supporting Information files are available without a subscription to ACS Web Editions. Such files may be downloaded by article for research use (if there is a public use license linked to the relevant article, that license may permit other uses). Permission may be obtained from ACS for other uses through requests via the RightsLink permission system: http://pubs.acs.org/page/copyright/permissions.html.

PDF [ 2MB]

Pair your accounts.

Export articles to Mendeley

Get article recommendations from ACS based on references in your Mendeley library.

Pair your accounts.

Export articles to Mendeley

Get article recommendations from ACS based on references in your Mendeley library.

You’ve supercharged your research process with ACS and Mendeley!

STEP 1:
Click to create an ACS ID

Please note: If you switch to a different device, you may be asked to login again with only your ACS ID.

Please note: If you switch to a different device, you may be asked to login again with only your ACS ID.

Please note: If you switch to a different device, you may be asked to login again with only your ACS ID.

MENDELEY PAIRING EXPIRED
Your Mendeley pairing has expired. Please reconnect