Myostatin gene knockout mediated by Cas9-D10A nickase in chicken DF1 cells without off-target effect.

ABSTRACT:

Objective

Based on rapid advancement of genetic modification techniques, genomic editing is expected to become the most efficient tool for improvement of economic traits in livestock as well as poultry. In this study, we examined and verified the nickase of mutated CRISPR-associated protein 9 (Cas9) to modulate the specific target gene in chicken DF1 cells.

Methods

Chicken myostatin which inhibits muscle cell growth and differentiation during myogenesis was targeted to be deleted and mutated by the Cas9-D10A nickase. After co-transfection of the nickase expression vector with green fluorescent gene (GFP) gene and targeted multiplex guide RNAs (gRNAs), the GFP-positive cells were sorted out by fluorescence-activated cell sorting procedure.

Results

Through the ge

SUBMITTER: Lee JH 

PROVIDER: S-EPMC5411835 | biostudies-literature | 2017 May

REPOSITORIES: biostudies-literature