Chemical Control of CRISPR Gene Editing via Conditional Diacylation Crosslinking of Guide RNAs. Issue 10 (3rd February 2023) - Lei, Huajun
Abstract: Conditional control of RNA structure and function has emerged as an effective toolkit. Here, a strategy based on a one‐step introduction of diacylation linkers and azide groups on the 2′‐OH of RNA is advance. Selected from eight phosphine reagents, it is found that 2‐(diphenylphosphino)ethylamine has excellent performance in reducing azides via a Staudinger reduction to obtain the original RNA. It is demonstrated that the enzymatic activities of Cas13 and Cas9 can be regulated by chemically modified guide RNAs, and further achieved ligand‐induced gene editing in living cells by a controllable CRISPR/Cas9 system. Abstract : In the current study, a diacylation crosslinking strategy is developed to regulate CRISPR systems and DPPEA is used to restore the RNA structure to conditionally control gene editing when needed.