Development of drug-inducible CRISPR-Cas9 systems for large-scale functional screening.

ABSTRACT:

Background

Large-scale genetic screening using CRISPR-Cas9 technology has emerged as a powerful approach to uncover and validate gene functions. The ability to control the timing of genetic perturbation during CRISPR screens will facilitate precise dissection of dynamic and complex biological processes. Here, we report the optimization of a drug-inducible CRISPR-Cas9 system that allows high-throughput gene interrogation with a temporal control.

Results

We designed multiple drug-inducible sgRNA expression vectors and measured their activities using an EGFP gene disruption assay in 11 human and mouse cell lines. The optimal design allows for a tight and inducible control of gene knockout in vitro, and in vivo during a seven-week-long experiment following hematopoietic reconsti

SUBMITTER: Sun N 

PROVIDER: S-EPMC6425629 | biostudies-literature | 2019 Mar

REPOSITORIES: biostudies-literature