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

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

Research output: Contribution to journalArticle

60 Citations (Scopus)

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

Fingerprint

Clustered Regularly Interspaced Short Palindromic Repeats
Drosophila
germ cells
engineering
genomics
Endonucleases
genome
Guide RNA
Muscidae
Genome
DNA Cleavage
genetically modified organisms
RNA
toxicity
mutation
Diptera
DNA
Embryonic Structures
Mutation

Keywords

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

ASJC Scopus subject areas

  • Insect Science
  • Medicine(all)

Cite this

A simplified and efficient germline-specific CRISPR/Cas9 system for Drosophila genomic engineering. / Sebo, Zachary L.; Lee, Han B.; Peng, Ying; Guo, Yi D.

In: Fly, Vol. 8, No. 1, 2014.

Research output: Contribution to journalArticle

Sebo, Zachary L. ; Lee, Han B. ; Peng, Ying ; Guo, Yi D. / A simplified and efficient germline-specific CRISPR/Cas9 system for Drosophila genomic engineering. In: Fly. 2014 ; Vol. 8, No. 1.
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