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

Here, we present a simple and highly efficient method for generating and detecting mutations ofany gene in Drosophila melanogaster through theuse of the CRISPR/Cas9 system (clustered regularlyinterspaced 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. © 2013 The Authors.