title: CRISPR/Cas9 mediated genome engineering in Drosophila.
abstract: Genome engineering has revolutionised genetic analysis in many organisms. Here we describe a simple and efficient technique to generate and detect novel mutations in desired target genes in Drosophila melanogaster. We target double strand breaks to specific sites within the genome by injecting mRNA encoding the Cas9 endonuclease and in vitro transcribed synthetic guide RNA into Drosophila embryos. The small insertion and deletion mutations that result from inefficient non-homologous end joining at this site are detected by high resolution melt analysis of whole flies and individual wings, allowing stable lines to be made within 1 month.
authors.affiliation: Medical Research Council Functional Genomics Unit, Department of Physiology, Anatomy and Genetics, University of Oxford, South Parks Road, Oxford OX1 3PT, United Kingdom. Electronic address: andrew.bassett@dpag.ox.ac.uk.
authors.forename: Andrew
authors.initials: A
authors.lastname: Bassett
authors.affiliation: Medical Research Council Functional Genomics Unit, Department of Physiology, Anatomy and Genetics, University of Oxford, South Parks Road, Oxford OX1 3PT, United Kingdom. Electronic address: jilong.liu@dpag.ox.ac.uk.
authors.forename: Ji-Long
authors.initials: JL
authors.lastname: Liu
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D001483:Base Sequence; D001665:Binding Sites / Q000502:physiology; D064113:CRISPR-Cas Systems / Q000235:genetics*; D064112:Clustered Regularly Interspaced Short Palindromic Repeats / Q000235:genetics*; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D004330:Drosophila; D029721:Drosophila Proteins / Q000235:genetics* / Q000378:metabolism; D005818:Genetic Engineering / Q000379:methods*; D016678:Genome / Q000235:genetics*; D008969:Molecular Sequence Data
chemical_list: D004268:DNA-Binding Proteins; D029721:Drosophila Proteins; C110593:cas protein, Drosophila
keywords: CRISPR; Cas9; Drosophila melanogaster; Genome engineering; RNA injection; Targeted mutagenesis