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

Zachary L. Sebo, Han B. Lee, Ying Peng, Yi Guo

Research output: Contribution to journalArticlepeer-review

64 Scopus citations

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.

Original languageEnglish (US)
JournalFly
Volume8
Issue number1
DOIs
StatePublished - 2014

Keywords

  • CRISPR/Cas9
  • Engineered endonuclease
  • Genomic engineering
  • Germline
  • RNA-guided DNA cleavage

ASJC Scopus subject areas

  • Insect Science

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