通过将 CRISPR/Cpf1 与光遗传学和还原响应基序相结合,我们开发了一种可光激活的交联 crRNA,可在光照下实现精确的基因组编辑。该系统还允许通过外部应用还原剂来终止编辑活动。双刺激响应 CRISPR/Cpf1 编辑过程以独特的 OFF → ON → OFF 顺序运行,使其成为研究时间敏感的生物事件的宝贵工具。
申请人:The Board of Trustees of the Leland Stanford Junior University
公开号:US20190264205A1
公开(公告)日:2019-08-29
Reagents and methods to cloak and uncloak RNA polymers and applications thereof are provided. Photocloaking molecules are used to label RNA polymers. Radiant energy is used to remove photoreleaseable protecting adducts and revert a RNA polymer to its native form.
rely on fully synthetic nucleic acids with photocaged nucleobases, limiting application to relatively short synthetic RNAs. Here we report a method to gain photocontrol over RNA by postsynthetic acylation of 2'-hydroxyls with photoprotecting groups. One-step introduction of these groups efficiently blocks hybridization, which is restored after light exposure. Polyacylation (termed cloaking) enables
Chemical control of CRISPR/Cpf1 editing <i>via</i> orthogonal activation and deactivation of crosslinked crRNA
作者:Cui-Lian Lin、Wen-Da Chen、Li Liu、Liang Cheng
DOI:10.1039/d4cc01106f
日期:——
Through the integration of CRISPR/Cpf1 with optogenetics and a reduction-responsive motif, we have developed a photoactivatable cross-linked crRNA that enables precise genome editing upon light exposure. This system also allows for termination of editing activity through external application of reducing agent. The dual-stimuli-responsive CRISPR/Cpf1 editing process operates in a unique OFF → ON → OFF
通过将 CRISPR/Cpf1 与光遗传学和还原响应基序相结合,我们开发了一种可光激活的交联 crRNA,可在光照下实现精确的基因组编辑。该系统还允许通过外部应用还原剂来终止编辑活动。双刺激响应 CRISPR/Cpf1 编辑过程以独特的 OFF → ON → OFF 顺序运行,使其成为研究时间敏感的生物事件的宝贵工具。