Highly Efficient Targeted Mutagenesis of Drosophila With the CRISPR/Cas9 System

Cell Rep. 2013 Jul 11;4(1):220-8. doi: 10.1016/j.celrep.2013.06.020. Epub 2013 Jul 1.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

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

Substances

  • Endodeoxyribonucleases