RNA-guided genome editing in Drosophila with the purified Cas9 protein

G3 (Bethesda). 2014 May 28;4(7):1291-5. doi: 10.1534/g3.114.012179.

Abstract

We report a method for generating Drosophila germline mutants effectively via injection of the complex of the purified Cas9 protein, tracrRNA, and gene-specific crRNAs, which may reduce delayed mutations because of the transient activity of the Cas9 protein, combined with the simple mutation detection in GO founders by the T7E1 assay.

Keywords: Cas9 protein; Drosophila; RGEN.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • Clustered Regularly Interspaced Short Palindromic Repeats / genetics
  • DNA-Binding Proteins / genetics
  • Drosophila / genetics
  • Drosophila Proteins / genetics*
  • Drosophila Proteins / metabolism
  • Genome*
  • Germ-Line Mutation
  • RNA Editing
  • RNA, Guide, CRISPR-Cas Systems

Substances

  • DNA-Binding Proteins
  • Drosophila Proteins
  • e protein, Drosophila