Conditional control of RNA-guided nucleic acid cleavage and gene editing
Conditional control of RNA-guided nucleic acid cleavage and gene editing
复制标题
RNA引导的核酸切割和基因编辑的条件控制
DOI：
10.1038/s41467-019-13765-3
复制
发表时间：
2020-01-03
影响因子：
16.6
通讯作者：
Zhou, Xiang
中科院分区：
文献类型：
--
作者：
Wang, Shao-Ru;Wu, Ling-Yu;Zhou, Xiang
原核生物使用称为CRISPR的重复基因组元件（群集定期间隔短的腔粒重复序列）来破坏入侵的遗传分子。尽管CRISPR系统已被广泛用于DNA和RNA技术，但确实会发生某些不良影响。例如，组成性主动的CRISPR系统可能会导致某些脱离目标效应的风险。在这里，我们将指导RNA（GRNA）的合成后掩盖和化学激活引入控制CRISPR系统。使用了RNA结构探针（2-氮甲基二酸咪唑唑唑酯）。此外，我们完成了活细胞中基因编辑的条件控制。这项概念验证研究表明，GRNA作为化学生物学的多功能工具的化学激活潜力。
Prokaryotes use repetitive genomic elements termed CRISPR (clustered regularly interspaced short palindromic repeats) to destroy invading genetic molecules. Although CRISPR systems have been widely used in DNA and RNA technology, certain adverse effects do occur. For example, constitutively active CRISPR systems may lead to a certain risk of off-target effects. Here, we introduce post-synthetic masking and chemical activation of guide RNA (gRNA) to controlling CRISPR systems. An RNA structure profiling probe (2-azidomethylnicotinic acid imidazolide) is used. Moreover, we accomplish conditional control of gene editing in live cells. This proof-of-concept study demonstrates promising potential of chemical activation of gRNAs as a versatile tool for chemical biology.