An in-library ligation strategy and its application in CRISPR/Cas9 screening of high-order gRNA combinations.

ABSTRACT: Simultaneous targeting multiple genes is a big advantage of CRISPR (clustered regularly interspaced short palindromic repeats) genome editing but challenging to achieve in CRISPR screening. The crosstalk among genes or gene products is a common and fundamental mechanism to ensure cellular stability and functional diversity. However, the screening approach to map high-order gene combinations to the interesting phenotype is still lacking. Here, we developed a universal in-library ligation strategy and applied it to generate multiplexed CRISPR library, which could perturb four pre-designed targets in a cell. We conducted in vivo CRISPR screening for potential guide RNA (gRNA) combinations inducing anti-tumor immune responses. Simultaneously disturbing a combination of three checkpoints in CD8

SUBMITTER: Lu Z 

PROVIDER: S-EPMC9226518 | biostudies-literature | 2022 Jun

REPOSITORIES: biostudies-literature