A simplified and efficient germline-specific CRISPR/Cas9 system for Drosophila genomic engineering

作者：Sebo Zachary L; Lee Han B; Peng Ying^*; Guo Yi

来源：Fly, 2014, 8(1): 52-57.

DOI：10.4161/fly.26828

• RNA-guided DNA cleavage
• germline
• genomic engineering
• engineered endonuclease
• CRISPR/Cas9

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.

• 出版日期2014-1-1