Precise Genome Editing of Drosophila with CRISPR RNA-Guided Cas9

Methods Mol Biol. 2015;1311:335-48. doi: 10.1007/978-1-4939-2687-9_22.

Abstract

The readily programmable CRISPR-Cas9 system is transforming genome engineering. We and others have adapted the S. pyogenes CRISPR-Cas9 system to precisely engineer the Drosophila genome and demonstrated that these modifications are efficiently transmitted through the germline. Here we provide a detailed protocol for engineering small indels, defined deletions, and targeted insertion of exogenous DNA sequences within one month using a rapid DNA injection-based approach.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • CRISPR-Associated Proteins / metabolism*
  • CRISPR-Cas Systems / genetics*
  • DNA Repair
  • Drosophila / embryology
  • Drosophila / genetics
  • Embryo, Nonmammalian / metabolism
  • Gene Targeting
  • Genetic Engineering / methods*
  • Genome / genetics*
  • Homologous Recombination
  • Plasmids / genetics
  • RNA / genetics*

Substances

  • CRISPR-Associated Proteins
  • RNA