earticle

영어

In this study, we mutated SH3 domain containing ring finger 2 (SH3RF2) gene to investigate the biological functionality through CRISPR/CRISPR-associated protein 9 (Cas9)-mediated genome editing technology. SH3RF2 has been reported for one of the critical domestication genes in chickens and it expected that SH3RF2 could be closely related to growth improvement in chicken. However, its biofunctionality still remains to be elucidated. Firstly, we cloned the partial genomic structure and identified sequences of the quail SH3RF2 gene. Subsequently, SH3RF2 was knocked out using CRISPR/Cas9 technology and single cell-derived SH3RF2 mutant line was established in quail myoblast (QM7) cells. As a result, we established an SH3RF2 knockout QM7#4 subline which had 61 and 155 nucleotide deletion mutations. After the induction of myotube differentiation, the expression profiles during muscle differentiation were analyzed and compared between regular QM7 and SH3RF2 KO QM7#4 cells by global RNA sequencing and bioinformatics analysis. Additionally, we also mutated SH3RF2 gene in chicken primordial germ cells (PGCs) to generate the SH3RF2 knockout chickens. Thus, to investigate the biofunctional activity of a predicted target gene, in vivo and in vitro functional genomic analyses should be a prerequisite.