Conditional control of RNA-guided nucleic acid cleavage and gene editing

• Published in: Nature communications Vol. 11; no. 1; pp. 91 - 10
• Main Authors: Wang, Shao-Ru, Wu, Ling-Yu, Huang, Hai-Yan, Xiong, Wei, Liu, Jian, Wei, Lai, Yin, Ping, Tian, Tian, Zhou, Xiang
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 03.01.2020; Nature Publishing Group; Nature Portfolio
• Subjects: 631/337/4041/3196; 631/92/610; 9/10; Activation; CRISPR; CRISPR-Cas Systems; Deoxyribonucleic acid; DNA; Endonucleases - metabolism; Gene Editing; Genetic modification; Genome editing; gRNA; HeLa Cells; Humanities and Social Sciences; Humans; Masking; multidisciplinary; Nucleic acids; Organic chemistry; Prokaryotes; Ribonucleic acid; RNA; RNA - genetics; RNA - metabolism; RNA probes; RNA, Guide, CRISPR-Cas Systems - genetics; RNA, Guide, CRISPR-Cas Systems - metabolism; Science; Science (multidisciplinary)
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-019-13765-3

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. Constituitively active CRISPR systems have the risk of adverse off-target effects. Here the authors use chemical masking and activation of gRNA to control activity.