KrCRISPR: an easy and efficient strategy for generating conditional knockout of essential genes in cells.

ABSTRACT:

Background

CRISPR/Cas9 system is a powerful tool for knocking out genes in cells. However, genes essential for cell survival cannot be directly knocked out. Traditionally, generation of conditional knockout cells requires multiple steps.

Results

In this study, we developed an easy and efficient strategy to generate conditional knockout cells by using double episomal vectors - one which expresses gRNA and Cas9 nuclease, and the other expresses an inducible rescue gene. Using this system which we named "krCRISPR" (knockout-rescue CRISPR), we showed that essential genes, HDAC3 and DNMT1, can be efficiently knocked out. When cells reach a desired confluency, the exogenous rescue genes can be silenced by the addition of doxycycline. Furthermore, the krCRISPR system

SUBMITTER: Wang B 

PROVIDER: S-EPMC6480908 | biostudies-literature | 2019

REPOSITORIES: biostudies-literature