title: A simplified and efficient germline-specific CRISPR/Cas9 system for Drosophila genomic engineering.
abstract: The type II CRISPR/Cas9 system (clustered regularly interspaced short palindromic repeats/CRISPR-associated) has recently emerged as an efficient and simple tool for site-specific engineering of eukaryotic genomes. To improve its applications in Drosophila genome engineering, we simplified the standard two-component CRISPR/Cas9 system by generating a stable transgenic fly line expressing the Cas9 endonuclease in the germline (Vasa-Cas9 line). By injecting vectors expressing engineered target-specific guide RNAs into Vasa-Cas9 fly embryos, mutations were generated from site-specific DNA cleavages and efficiently transmitted into progenies. Because Cas9 endonuclease is the universal component of the type II CRISPR/Cas9 system, site-specific genomic engineering based on this improved platform can be achieved with lower complexity and toxicity, greater consistency, and excellent versatility.
authors.affiliation: Department of Biochemistry and Molecular Biology; Mayo Clinic; Rochester, MN USA; Division of Molecular Biology and Biochemistry; University of Missouri-Kansas City; Kansas City, MO USA.
authors.forename: Zachary L
authors.initials: ZL
authors.lastname: Sebo
authors.affiliation: Graduate Program in Neurobiology of Disease; Mayo Graduate School; Mayo Clinic; Rochester, MN USA.
authors.forename: Han B
authors.initials: HB
authors.lastname: Lee
authors.affiliation: Department of Biochemistry and Molecular Biology; Mayo Clinic; Rochester, MN USA.
authors.forename: Ying
authors.initials: Y
authors.lastname: Peng
authors.affiliation: Department of Biochemistry and Molecular Biology; Mayo Clinic; Rochester, MN USA; Division of Gastroenterology and Hepatology; Mayo Clinic; Rochester, MN USA.
authors.forename: Yi
authors.initials: Y
authors.lastname: Guo
mesh_terms: D000818:Animals; D064112:Clustered Regularly Interspaced Short Palindromic Repeats; D004330:Drosophila / Q000235:genetics*; D004720:Endonucleases / Q000235:genetics*; D005260:Female; D005818:Genetic Engineering; D008297:Male
chemical_list: D004720:Endonucleases
keywords: CRISPR/Cas9; RNA-guided DNA cleavage; engineered endonuclease; genomic engineering; germline