title: Highly efficient targeted mutagenesis of Drosophila with the CRISPR/Cas9 system.
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.
authors.affiliation: Medical Research Council Functional Genomics Unit, Department of Physiology, Anatomy and Genetics, University of Oxford, South Parks Road, Oxford, OX1 3PT, UK. andrew.bassett@dpag.ox.ac.uk
authors.forename: Andrew R
authors.initials: AR
authors.lastname: Bassett
authors.forename: Charlotte
authors.initials: C
authors.lastname: Tibbit
authors.forename: Chris P
authors.initials: CP
authors.lastname: Ponting
authors.forename: Ji-Long
authors.initials: JL
authors.lastname: Liu
mesh_terms: D000818:Animals; D001483:Base Sequence; D064112:Clustered Regularly Interspaced Short Palindromic Repeats / Q000235:genetics*; D004330:Drosophila / Q000235:genetics*; D004706:Endodeoxyribonucleases / Q000235:genetics* / Q000378:metabolism; D018095:Germ-Line Mutation; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed / Q000379:methods*
chemical_list: D004706:Endodeoxyribonucleases