RNA-guided genome editing of mammalian cells

Methods Mol Biol. 2014:1114:269-77. doi: 10.1007/978-1-62703-761-7_17.

Abstract

The microbial CRISPR-Cas adaptive immune system can be harnessed to facilitate genome editing in eukaryotic cells (Cong L et al., Science 339, 819-823, 2013; Mali P et al., Science 339, 823-826, 2013). Here we describe a protocol for the use of the RNA-guided Cas9 nuclease from the Streptococcus pyogenes type II CRISPR system to achieve specific, scalable, and cost-efficient genome editing in mammalian cells.

Publication types

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

MeSH terms

  • Animals
  • Cell Line
  • Cloning, Molecular
  • DNA End-Joining Repair
  • Gene Order
  • Gene Targeting / methods
  • Genetic Vectors
  • Genome*
  • Humans
  • RNA Editing*
  • RNA, Small Untranslated
  • Transfection

Substances

  • RNA, Small Untranslated