Efficient Gene Knock-out and Knock-in with Transgenic Cas9 in Drosophila. Issue 5 (1st May 2014)

• Record Type: Journal Article
• Authors: Xue, Zhaoyu; Ren, Mengda; Wu, Menghua; Dai, Junbiao; Rong, Yikang S; Gao, Guanjun
• Is Part Of: G3. Volume 4:Issue 5(2014)
• Page Start: 925
• Page End: 929
• Publication Date: 2014-05-01
• Subjects: knock-out -- knock-in -- Cas9 -- gRNA -- Drosophila; Genetics -- Research -- Periodicals; Genomics -- Periodicals; Genetics; Genomics; Genes; Genetics -- Research; Genomics; Electronic journals; Periodical; Periodicals; Fulltext; Internet Resources; Periodicals; 572.8
• DOI: 10.1534/g3.114.010496
• Languages: English
• ISSNs: 2160-1836

Abstract

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.