Imperial College London
Alternate

Professor Molly Stevens

Faculty of Engineering, Department of Materials

Professor of Biomedical Materials and Regenerative Medicine
 
 
 
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Contact

 

+44 (0)20 7594 6804m.stevens

 
 
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Location

 

208Royal School of MinesSouth Kensington Campus

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Summary

 

Publications

Citation

BibTex format

@article{Moroz-Omori:2020:10.1021/acscentsci.9b01093,
author = {Moroz-Omori, EV and Satyapertiwi, D and Ramel, MC and Hogset, H and Sunyovszki, I and Liu, Z and Wojciechowski, J and Zhang, Y and Grigsby, CL and Casimiro, Brito L and Bugeon, L and Dallman, M and Stevens, M},
doi = {10.1021/acscentsci.9b01093},
journal = {ACS Central Science},
pages = {695--703},
title = {Photoswitchable gRNAs for spatiotemporally controlled CRISPR-Cas-based genomic regulation},
url = {http://dx.doi.org/10.1021/acscentsci.9b01093},
volume = {6},
year = {2020}
}
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RIS format (EndNote, RefMan)

TY  - JOUR
AB  - The recently discovered CRISPR-Cas gene editing system and its derivatives have found numerous applications in fundamental biology research and pharmaceutical sciences. The need for precise external control over the gene editing and regulatory events has driven the development of inducible CRISPR-Cas systems. While most of the light-controllable CRISPR-Cas systems are based on protein engineering, we developed an alternative synthetic approach based on modification of crRNA/tracrRNA duplex (guide RNA or gRNA) with photocaging groups, preventing the gRNA from recognizing its genome target sequence until its deprotection is induced within seconds of illumination. This approach relies on a straightforward solid-phase synthesis of the photocaged gRNAs, with simpler purification and characterization processes in comparison to engineering a light-responsive protein. We have demonstrated the feasibility of photocaging of gRNAs and light-mediated DNA cleavage upon brief exposure to light in vitro. We have achieved light-mediated spatiotemporally resolved gene editing as well as gene activation in cells, whereas photocaged gRNAs showed virtually no detectable gene editing or activation in the absence of light irradiation. Finally, we have applied this system to spatiotemporally control gene editing in zebrafish embryos in vivo, enabling the use of this strategy for developmental biology and tissue engineering applications.
AU  - Moroz-Omori,EV
AU  - Satyapertiwi,D
AU  - Ramel,MC
AU  - Hogset,H
AU  - Sunyovszki,I
AU  - Liu,Z
AU  - Wojciechowski,J
AU  - Zhang,Y
AU  - Grigsby,CL
AU  - Casimiro,Brito L
AU  - Bugeon,L
AU  - Dallman,M
AU  - Stevens,M
DO  - 10.1021/acscentsci.9b01093
EP  - 703
PY  - 2020///
SN  - 2374-7943
SP  - 695
TI  - Photoswitchable gRNAs for spatiotemporally controlled CRISPR-Cas-based genomic regulation
T2  - ACS Central Science
UR  - http://dx.doi.org/10.1021/acscentsci.9b01093
UR  - http://hdl.handle.net/10044/1/79025
VL  - 6
ER  -
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