학술논문

Highly Efficient Targeted Mutagenesis of Drosophila with the CRISPR/Cas9 System
Document Type
article
Source
Cell Reports, Vol 4, Iss 1, Pp 220-228 (2013)
Subject
Biology (General)
QH301-705.5
Language
English
ISSN
2211-1247
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.