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

Fly (Austin). 2014;8(1):52-7. doi: 10.4161/fly.26828. Epub 2013 Oct 18.


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.

Keywords: CRISPR/Cas9; RNA-guided DNA cleavage; engineered endonuclease; genomic engineering; germline.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Clustered Regularly Interspaced Short Palindromic Repeats*
  • Drosophila / genetics*
  • Endonucleases / genetics*
  • Female
  • Genetic Engineering*
  • Male


  • Endonucleases