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Spatial and Temporal Control of CRISPR-Cas9-Mediated Gene Editing Delivered via a Light-Triggered Liposome System

  • Yagiz Alp Aksoy
    Yagiz Alp Aksoy
    Diabetes and Metabolism Division, Garvan Institute of Medical Research, Sydney, NSW 2010, Australia
    Department of Biomedical Sciences, Faculty of Medicine and Health Sciences, Centre for Motor Neuron Disease Research, Macquarie University, Sydney, NSW 2109, Australia
    Sydney Medical School, The University of Sydney, Sydney, NSW 2006, Australia
  • Biyao Yang
    Biyao Yang
    ARC Centre of Excellence for Nanoscale Biophotonics, Graduate School of Biomedical Engineering, University of New South Wales, Sydney, NSW 2052, Australia
    More by Biyao Yang
  • Wenjie Chen
    Wenjie Chen
    Center for Pharmaceutical Engineering and Sciences, Department of Pharmaceutics, School of Pharmacy, Virginia Commonwealth University, Richmond, Virginia 23298, United States
    More by Wenjie Chen
  • Tzongtyng Hung
    Tzongtyng Hung
    The Biological Resource Imaging Laboratory, University of New South Wales, Sydney, NSW 2052, Australia
  • Rhiannon P. Kuchel
    Rhiannon P. Kuchel
    Electron Microscope Unit, University of New South Wales, Sydney, NSW 2052, Australia
  • Nathan W. Zammit
    Nathan W. Zammit
    Garvan Institute of Medical Research, Sydney, NSW 2010, Australia
    Faculty of Medicine, University of New South Wales, Sydney, NSW 2052, Australia
  • Shane T. Grey
    Shane T. Grey
    Garvan Institute of Medical Research, Sydney, NSW 2010, Australia
    Faculty of Medicine, University of New South Wales, Sydney, NSW 2052, Australia
  • Ewa M. Goldys*
    Ewa M. Goldys
    ARC Centre of Excellence for Nanoscale Biophotonics, Graduate School of Biomedical Engineering, University of New South Wales, Sydney, NSW 2052, Australia
    *Email: [email protected]
  • , and 
  • Wei Deng*
    Wei Deng
    ARC Centre of Excellence for Nanoscale Biophotonics, Graduate School of Biomedical Engineering, University of New South Wales, Sydney, NSW 2052, Australia
    *Email: [email protected]
    More by Wei Deng
Cite this: ACS Appl. Mater. Interfaces 2020, 12, 47, 52433–52444
Publication Date (Web):November 11, 2020
https://doi.org/10.1021/acsami.0c16380
Copyright © 2020 American Chemical Society

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    Abstract

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    The CRISPR-Cas9 and related systems offer a unique genome-editing tool allowing facile and efficient introduction of heritable and locus-specific sequence modifications in the genome. Despite its molecular precision, temporal and spatial control of gene editing with the CRISPR-Cas9 system is very limited. We developed a light-sensitive liposome delivery system that offers a high degree of spatial and temporal control of gene editing with the CRISPR-Cas9 system. We demonstrated its efficient protein release by respectively assessing the targeted knockout of the eGFP gene in human HEK293/GFP cells and the TNFAIP3 gene in TNFα-induced HEK293 cells. We further validated our results at a single-cell resolution using an in vivo eGFP reporter system in zebrafish (77% knockout). These findings indicate that light-triggered liposomes may have new options for precise control of CRISPR-Cas9 release and editing.

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    The Supporting Information is available free of charge at https://pubs.acs.org/doi/10.1021/acsami.0c16380.

    • Experimental setup of the cell work, cellular uptake activity of the liposomes, TEM images of the liposomes loaded with the Cas9 and sgRNA complex, serum stability of the liposomes, cell’s viability assessment, 3D rendered confocal images of individual slow-muscle fibers expressing eGFP, and quantitative assessment in zebrafish by counting the number of knockout slow-muscle fibers per embryo (PDF)

    • Fluorescent signal of eGFP across individual slow-twitch muscle fibers (Video S1) (MP4) (MOV)

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