Photoswitchable gRNAs for Spatiotemporally ControlledCRISPR-Cas-Based Genomic Regulation

This is the authors' abstract. We don't add key points for this paper.

The recently discovered-Cas gene editing system and its have found numerous applications in fundamental biology and pharmaceutical sciences. The need for precise external over the gene editing and regulatory events has driven the of inducible CRISPR-Cas systems. While most of the light-controllable-Cas systems are based on protein engineering, we developed alternative synthetic approach based on modification of crRNA/tracrRNA (guide RNA or gRNA) with photocaging groups, preventing the from recognizing its genome target sequence until its deprotection induced within seconds of illumination. This approach relies on straightforward solid-phase synthesis of the photocaged gRNAs, with purification and characterization processes in comparison engineering a light-responsive protein. We have demonstrated the of photocaging of gRNAs and light-mediated DNA cleavage brief exposure to light in vitro. We have achieved-mediated spatiotemporally resolved gene editing as well as gene in cells, whereas photocaged gRNAs showed virtually no gene editing or activation in the absence of light irradiation., we have applied this system to spatiotemporally control gene in zebrafish embryos in vivo, enabling the of this strategy for developmental biology and tissue engineering.