Very fast CRISPR on demand
A caged RNA strategy that allows Cas9 to bind DNA but not cleave until light-induced activation is developed, referred to as very fast CRISPR (vfCRISPR), which enables DNA repair studies at high resolution in space and time at specific genome locations.
Abstract
<jats:title>Very fast CRISPR on demand</jats:title> <jats:p> Numerous efforts have been made to improve the temporal resolution of CRISPR-Cas9–mediated DNA cleavage to the hour time scale. Liu <jats:italic>et al.</jats:italic> developed a Cas9 system that achieved genome-editing manipulation at the second time scale (see the Perspective by Medhi and Jasin). Part of the guide RNA is chemically caged, allowing the Cas9-guide RNA complex to bind at a specific genomic locus without cleavage until activation by light. This fast CRISPR system achieves genome editing at high temporal resolution, enabling the study of early molecular events of DNA repair processes. This system also has high spatial resolution at short time scales, allowing editing of one genomic allele while leaving the other unperturbed. </jats:p> <jats:p> <jats:italic>Science</jats:italic> , this issue p. <jats:related-article xmlns:xlink="http://www.w3.org/1999/xlink" ext-link-type="doi" issue="6496" page="1265" related-article-type="in-this-issue" vol="368" xlink:href="10.1126/science.aay8204">1265</jats:related-article> ; see also p. <jats:related-article xmlns:xlink="http://www.w3.org/1999/xlink" ext-link-type="doi" issue="6496" page="1180" related-article-type="in-this-issue" vol="368" xlink:href="10.1126/science.abc3997">1180</jats:related-article> </jats:p>