Optimal LentiCRISPR-Based System for Sequential CRISPR/Cas9 Screens.

ABSTRACT: The advent of genome-wide clustered regularly interspaced short palindromic repeats (CRISPR) screening has advanced the understanding of molecular systems within cells. Here, we demonstrate the utility of sequentially performed CRISPR knockout screens that use an existing library to explore a biological question across the human genome, and then the remaining cells are used to examine each gene candidate against one common gene of interest. We call this approach "Many vs One" CRISPR screening, made possible by a modified 7SK promoter in place of the U6 promoter to drive expression of a single guide RNA. Inserting this novel 7SK promoter into the ubiquitously used lentiCRISPRv2 backbone is crucial, because it overcomes the need for a substantial increase in CRISPR library coverage during sc

SUBMITTER: Hutcheson RL 

PROVIDER: S-EPMC9802680 | biostudies-literature | 2022 Jul

REPOSITORIES: biostudies-literature