Photocontrol of CRISPR/Cas9 function by site-specific chemical modification of guide RNA.

ABSTRACT: The function of CRISPR/Cas9 can be conditionally controlled by the rational engineering of guide RNA (gRNA) to target the gene of choice for precise manipulation of the genome. Particularly, chemically modified gRNA that can be activated by using specific stimuli provides a unique tool to expand the versatility of conditional control. Herein, unlike previous engineering of gRNA that generally focused on the RNA part only but neglected RNA-protein interactions, we aimed at the interactive sites between 2'-OH of ribose in the seed region of gRNA and the Cas9 protein and identified that chemical modifications at specific sites could be utilized to regulate the Cas9 activity. By introducing a photolabile group at these specific sites, we achieved optical control of Cas9 activity without disrup

SUBMITTER: Wang Y 

PROVIDER: S-EPMC8162494 | biostudies-literature | 2020 Sep

REPOSITORIES: biostudies-literature