ZFIN ID: ZDB-PUB-200403-35
Spatiotemporal Control of CRISPR/Cas9 Function in Cells and Zebrafish using Light-Activated Guide RNA
Zhou, W., Brown, W., Bardhan, A., Delaney, M., Ilk, A., Rauen, R., Kahn, S., Tsang, M., Deiters, A.
Date: 2020
Source: Angewandte Chemie (International ed. in English)   59(23): 8998-9003 (Journal)
Registered Authors: Tsang, Michael
Keywords: CRISPR/Cas9, caged compounds, gene editing, optical control, zebrafish
MeSH Terms:
  • Animals
  • CRISPR-Cas Systems/genetics*
  • Cell Line
  • Gene Editing/methods*
  • Nucleic Acid Hybridization
  • RNA, Guide/genetics*
  • Spatio-Temporal Analysis
  • Time Factors
  • Zebrafish*
PubMed: 32160370 Full text @ Angew. Chem. Int. Ed. Engl.
ABSTRACT
We developed a new method for conditional regulation of CRISPR/Cas9 activity in mammalian cells and zebrafish embryos via photochemically activated, caged guide RNAs. Caged gRNAs are generated by substituting four nucleobases evenly distributed throughout the 5'-protospacer region with caged nucleobases during synthesis. Caging confers complete suppression of gRNA:dsDNA- target hybridization and rapid restoration of CRISPR/Cas9 function upon optical activation. This tool offers simplicity and complete programmability in design, high spatiotemporal specificity in cells and zebrafish embryos, excellent off to on switching, and stability by preserving the ability to form Cas9:gRNA ribonucleoprotein complexes. Caged gRNAs are novel tools for conditional control of gene editing, thereby enabling the investigation of spatiotemporally complex physiological events by obtaining a better understanding of dynamic gene regulation.
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