Journal Club-Genome editing with RNA guided Cas9 nuclease in Zebrafish embryos

This is an original article published in early 2013 in Cell Research magazine.

It mainly includes four parts of experiments: Cas9/gRNA ribonucleoprotein complex executes site-specific DNA cleavage in mammalian cells; Cas9/gRNA induces indels in the etsrp locus in zebrafish; Cas9/gRNA induces indels in the gata4and gata5 loci in zebrafish; Cas9/gRNA induces insertion of mloxP at the break site ofetsrp in zebrafish.

The researchers also use representative SURVEYOR assay, which shows higher efficiency of Cas9-mediated cleavage or indels than that by TALENs. Besides, their work presents two additional important findings: the biallelic knockouts by Cas9/gRNA and site-specific insertions of mloxP for the potential generation of conditional knockouts.

Compared with ZFN or TALEN, the Cas9/gRNA system is an easier and economical way to design and manufacture genome editing tools, and has comparable efficiency in generating site-specific indels.