Highly efficient targeted mutagenesis of Drosophila with the CRISPR/Cas9 system

Andrew R Bassett, Charlotte Tibbit, Chris P Ponting, Ji-Long Liu

Research output: Contribution to journalArticlepeer-review

Abstract

Here, we present a simple and highly efficient method for generating and detecting mutations of any gene in Drosophila melanogaster through the use of the CRISPR/Cas9 system (clustered regularly interspaced palindromic repeats/CRISPR-associated). We show that injection of RNA into the Drosophila embryo can induce highly efficient mutagenesis of desired target genes in up to 88% of injected flies. These mutations can be transmitted through the germline to make stable lines. Our system provides at least a 10-fold improvement in efficiency over previously published reports, enabling wider application of this technique. We also describe a simple and highly sensitive method of detecting mutations in the target gene by high-resolution melt analysis and discuss how the new technology enables the study of gene function.

Original languageEnglish
Pages (from-to)220-8
Number of pages9
JournalCell Reports
Volume4
Issue number1
DOIs
Publication statusPublished - 11 Jul 2013

Keywords

  • Animals
  • Base Sequence
  • Clustered Regularly Interspaced Short Palindromic Repeats
  • Drosophila
  • Endodeoxyribonucleases
  • Germ-Line Mutation
  • Molecular Sequence Data
  • Mutagenesis, Site-Directed

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