Highly efficient targeted mutagenesis of Drosophila with the CRISPR/Cas9 system

Journal article

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: Bassett, AR; Tibbit, C; Ponting, CP; Liu, J-L

• Publication status: Published
• Peer review status: Peer reviewed
• Publisher: Cell Press
• Journal: Cell Reports
• Volume: 4
• Issue: 1
• Pages: 220-228
• Publication date: 2013-07-01
• Acceptance date: 2013-06-20
• DOI: 10.1016/j.celrep.2013.06.020
• EISSN: 2211-1247
• Language: English