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• W4224291206 abstract "Abstract It is important to control CRISPR/Cas9 when sufficient editing is obtained. In the current study, rational engineering of guide RNAs (gRNAs) is performed to develop small-molecule-responsive CRISPR/Cas9. For our purpose, the sequence of gRNAs are modified to introduce ligand binding sites based on the rational design of ligand–RNA pairs. Using short target sequences, we demonstrate that the engineered RNA provides an excellent scaffold for binding small molecule ligands. Although the ‘stem–loop 1’ variants of gRNA induced variable cleavage activity for different target sequences, all ‘stem–loop 3’ variants are well tolerated for CRISPR/Cas9. We further demonstrate that this specific ligand–RNA interaction can be utilized for functional control of CRISPR/Cas9 in vitro and in human cells. Moreover, chemogenetic control of gene editing in human cells transfected with all-in-one plasmids encoding Cas9 and designer gRNAs is demonstrated. The strategy may become a general approach for generating switchable RNA or DNA for controlling other biological processes." @default.
• W4224291206 created "2022-04-26" @default.
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• W4224291206 date "2022-04-21" @default.
• W4224291206 modified "2023-10-14" @default.
• W4224291206 title "Rational guide RNA engineering for small-molecule control of CRISPR/Cas9 and gene editing" @default.
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• W4224291206 doi "https://doi.org/10.1093/nar/gkac255" @default.
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