Journal
CELL REPORTS
Volume 4, Issue 1, Pages 220-228Publisher
CELL PRESS
DOI: 10.1016/j.celrep.2013.06.020
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Funding
- European Research Council (DARCGENs) [249869]
- Medical Research Council
- MRC [MC_U137788471, MC_UU_12021/1, MC_U137761446, MC_UU_12021/3] Funding Source: UKRI
- Medical Research Council [MC_UU_12021/1, MC_U137788471, MC_U137761446, MC_UU_12021/3] Funding Source: researchfish
- European Research Council (ERC) [249869] Funding Source: European Research Council (ERC)
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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.
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