2-(Azidomethyl)nicotinic acid imidazolide (NAI-N3) is an RNA icSHAPE probe for live-cell RNA structure profiling across the genome. icSHAPE -- or in vivo click selective 2′-hydroxyl acylation and profiling experiment -- uses NAI-N3 in a chemoaffinity method to probe RNA structure. NAI-N3 is an azido version of the cell-permeable SHAPE reagent 2-methylnicotinic acid imidazolide (NAI) that permits the tagging of NAI-N3-modified RNA with a biotin moiety for subsequent capture via streptavidin. Not only do such strategies further the understanding of RNA structure in living cells but also provide a tool for identifying regions that may be susceptible to therapeutic targeting. Recently, the azidomethylnicotinyl (AMN) group of NAI-N3 was demonstrated to block the function of gRNA and CRISPR systems, which could be reactivated by removing the AMN groups with Staudinger reduction (DPBM phosphine), overall providing a means to control nucleic acid cleavage and gene editing in live cells.
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
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