Generation of dual-gRNA library for combinatorial CRISPR screening of synthetic lethal gene pairs.

ABSTRACT: Combinatorial CRISPR screening is useful for investigating synthetic lethality (SL) gene pairs. Here, we detail the steps for dual-gRNA library construction, with the introduction of two backbones, LentiGuide_DKO and LentiCRISPR_DKO. We describe steps for in vitro screening with 22Rv1-Cas9 and SaOS2-Cas9 cells followed by sequencing and data analysis. By introducing two backbones, we optimized the library construction process, facilitated standard pair-end sequencing, and provided options of screening on cells with or without modification of Cas9 expression.

SUBMITTER: Tang S 

PROVIDER: S-EPMC9428847 | biostudies-literature | 2022 Sep

REPOSITORIES: biostudies-literature