alexa Highly efficient targeted mutagenesis of Drosophila with the CRISPR Cas9 system.
Medicine

Medicine

Advanced Techniques in Biology & Medicine

Author(s): Bassett AR, Tibbit C, Ponting CP, Liu JL

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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. Copyright © 2013 The Authors. Published by Elsevier Inc. All rights reserved.
This article was published in Cell Rep and referenced in Advanced Techniques in Biology & Medicine

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