Abstract
CRISPR (clustered regularly interspaced short palindromic repeats) systems have been established as valuable genome‐editing tools. Controlling CRISPR systems has high biological significance and this field has garnered intense interest. There is a considerable need for simple approaches with no need for protein engineering. The CRISPR systems usually require a guide RNA (gRNA) moiety to recruit and direct the nuclease complexes. In this respect, the ninhydrin (1,2,3‐indantrione monohydrate) seems to have considerable potential, as yet unexploited, for modifying gRNA. In this study, ninhydrin chemistry is explored for reversible postsynthetic modification of gRNA molecules. It is further shown that ninhydrin chemistry is efficient in modulating two important CRISPR systems. Thus, ninhydrin chemistry exhibits potential applications in future chemical biology studies.
Highlights
We further demonstrate that the ninhydrin-gRNA adducts can be readily reverted to original gRNAs under mild conditions
We further show that ninhydrin chemistry is efficient in manipulating CRISPR systems
In summary, we demonstrate the effectiveness of ninhydrin chemistry for conditional control of RNA-guided nucleic acid cleavage
Summary
The CRISPR (clustered regularly interspaced short palindromic repeats) system is a prevalent prokaryotic defense system against foreign genetic elements.[1,2,3,4] CRISPR systems have been established as valuable genome-editing tools.[5,6,7] The CRISPR system usually requires a guide RNA (gRNA) moiety to recruit and direct the CRISPR-associated proteins (Cas) ).[5,6,7] Controlling CRISPR systems has high biological significance and this field has garnered intense interests.[8,9,10,11] A large amount of attention has been directed especially to modifying the Cas proteins.[12,13,14,15,16,17] There is a considerable need for simpler approaches with no need for protein engineering. We intend to target gRNAs for compounds is much more electrophilic than that in a simple aldehyde or ketone.[29] In terms of structure, ninhydrin (1,2,3indantrione monohydrate) represents an ideal compound having three (potentially efficient in condensation reaction) carbonyl groups in one molecule.[30] Though ninhydrin is traditionally used to detect amino acids and amines, its reaction with heterocyclic nucleobases has been reported.[31,32] The site of reaction is believed to be at the amino groups of those nucleobases, which act as 1,3-binucleophiles In this respect, the ninhydrin seems to have considerable potential, as yet unexploited, for modifying gRNA molecules
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