Poster abstracts
Poster number 82 submitted by Wenyuan Zhou
Spatiotemporal control of CRISPR/Cas9 function in cells and zebrafish using light‐activated guide RNA
Wenyuan Zhou (Department of Chemistry, University of Pittsburgh), Wes Brown, Anirban Bardhan (Department of Chemistry, University of Pittsburgh), Michael Delaney, Amber S. Ilk, Randy R. Rauen, Shoeb I. Kahn (Horizon Discovery), Michael Tsang (Department of Developmental Biology, School of Medicine, University of Pittsburgh), Alexander Deiters (Department of Chemistry, University of Pittsburgh)
Abstract:
We developed a new method for the conditional regulation of CRISPR/Cas9 activity in mammalian cells and zebrafish embryos using photochemically activated, caged guide RNAs (gRNAs). 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 binding to target DNA 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 the conditional control of gene editing and transcriptional regulation, thereby enabling the investigation of spatiotemporally complex physiological events by obtaining a better understanding of dynamic gene regulation.
References:
Zhou, Wenyuan, et al. "Spatiotemporal Control of CRISPR/Cas9 Function in Cells and Zebrafish using Light‐Activated Guide RNA." Angewandte Chemie 132.23 (2020): 9083-9088.
Keywords: CRISPRCas9, optical control, RNA