Efficient gene disruption in cultured primary human endothelial cells by CRISPR/Cas9.

ABSTRACT:

Rationale

The participation of endothelial cells (EC) in many physiological and pathological processes is widely modeled using human EC cultures, but genetic manipulation of these untransformed cells has been technically challenging. Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 nuclease (Cas9) technology offers a promising new approach. However, mutagenized cultured cells require cloning to yield homogeneous populations, and the limited replicative lifespan of well-differentiated human EC presents a barrier for doing so.

Objective

To create a simple but highly efficient method using CRISPR/Cas9 to generate biallelic gene disruption in untransformed human EC.

Methods and results

To demonstrate proof-of-principle, we use

SUBMITTER: Abrahimi P 

PROVIDER: S-EPMC4490936 | biostudies-literature | 2015 Jul

REPOSITORIES: biostudies-literature