Journal
METHODS
Volume 69, Issue 2, Pages 128-136Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ymeth.2014.02.019
Keywords
Drosophila melanogaster; CRISPR; Cas9; Genome engineering; Targeted mutagenesis; RNA injection
Funding
- UK Medical Research Council
- European Research Council (DARCGENs) [249869]
- MRC [MC_UU_12021/3, MC_U137788471] Funding Source: UKRI
- Medical Research Council [MC_UU_12021/3, MC_U137788471] Funding Source: researchfish
- European Research Council (ERC) [249869] Funding Source: European Research Council (ERC)
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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. (C) 2014 Published by Elsevier Inc.
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