Xue, Z., Ren, M., Wu, M., Dai, J., Rong, Y.S., Gao, G. (2014). Efficient Gene Knock-out and Knock-in with Transgenic Cas9 in Drosophila. G3 (Bethesda) 4(5): 925--929.
FlyBase ID
FBrf0225034
Publication Type
Research paper
Abstract
Bacterial Cas9 nuclease induces site-specific DNA breaks using small gRNA as guides. Cas9 has been successfully introduced into Drosophila for genome editing. Here, we improve the versatility of this method by developing a transgenic system that expresses Cas9 in the Drosophila germline. Using this system, we induced inheritable knock-out mutations by injecting only the gRNA into embryos, achieved highly efficient mutagenesis by expressing gRNA from the promoter of a novel non-coding RNA gene, and recovered homologous recombination-based knock-in of a fluorescent marker at a rate of 4.5% by co-injecting gRNA with a circular DNA donor.
