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

Nature communications (2020-01-05)
Shao-Ru Wang, Ling-Yu Wu, Hai-Yan Huang, Wei Xiong, Jian Liu, Lai Wei, Ping Yin, Tian Tian, Xiang Zhou
RESUMEN

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.

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Sigma-Aldrich
2-Methylnicotinic acid imidazolide
Sigma-Aldrich
2-(Diphenylphosphino)benzoic acid, 97%
Sigma-Aldrich
2-(Azidomethyl)nicotinic acid imidazolide, ≥95%
Sigma-Aldrich
2-(Diphenylphosphanyl)benzamide, ≥95%