| Literature DB >> 24576617 |
Andrew Bassett1, Ji-Long Liu2.
Abstract
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.Entities:
Keywords: CRISPR; Cas9; Drosophila melanogaster; Genome engineering; RNA injection; Targeted mutagenesis
Mesh:
Substances:
Year: 2014 PMID: 24576617 DOI: 10.1016/j.ymeth.2014.02.019
Source DB: PubMed Journal: Methods ISSN: 1046-2023 Impact factor: 3.608