Cell Biology and Toxicology ( IF 5.3 ) Pub Date : 2022-05-30 , DOI: 10.1007/s10565-022-09723-3 Jizhong Zhao 1 , Hongmei Hu 1 , Hongling Zhou 1 , Jingwen Zhang 1 , Li Wang 2 , Rui Wang 1, 3
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We report development of a controllable gene editing tool that boronated gRNA, simply generated in situ, could regulate binding of gRNA molecules with either Cas9 endonuclease or target genes, thus serving as a modulator that can control CRISPR-Cas9 gene editing. Subsequent treatment with H2O2 facilitates the restoration of gene editing ability of the boronated gRNA to the level of using untreated gRNA. This is one of the few cases using small molecule to regulate CRISPR-Cas9 gene editing, which is a complement to the light approach, displaying great application potential.
Graphical abstract
We develop a controllable gene editing tools based on the CRISPR-Cas9 gene editing system. This tool can be regulated by oxidative small molecule, i.e., H2O2. Compared with the light method, the application scope of our CRISPR-Cas9 systems have been widened with the small-molecule-triggered approaches, preventing the potential damage of cells or organism caused by UV light. In addition, the gain-of-function tools are expanding the gene code expansion for mechanistic studies of target enzymes since it provides a positive route to evaluate the activity of a given enzyme in dynamic and inversible regulation of targeting cellular processes.
中文翻译：
活性氧信号分子诱导调控CRISPR-Cas9基因编辑
我们报告了一种可控基因编辑工具的开发，该工具可以简单地原位生成硼化 gRNA，从而调节 gRNA 分子与 Cas9 核酸内切酶或靶基因的结合，从而充当控制 CRISPR-Cas9 基因编辑的调节剂。随后用 H 2 O 2处理有助于将硼化 gRNA 的基因编辑能力恢复到使用未处理 gRNA 的水平。这是利用小分子调控CRISPR-Cas9基因编辑的少数案例之一，是对光方法的补充，显示出巨大的应用潜力。
图形概要
我们开发了基于CRISPR-Cas9基因编辑系统的可控基因编辑工具。该工具可以通过氧化小分子，即H 2 O 2来调节。与光方法相比，我们的CRISPR-Cas9系统通过小分子触发方法拓宽了应用范围，防止了紫外线对细胞或生物体造成的潜在损伤。此外，功能获得工具正在扩展用于目标酶机制研究的基因代码扩展，因为它提供了评估给定酶在靶向细胞过程的动态和可逆调节中的活性的积极途径。
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