Targeted Genome Engineering in Human Cells With the Cas9 RNA-guided Endonuclease

Nat Biotechnol. 2013 Mar;31(3):230-2. doi: 10.1038/nbt.2507. Epub 2013 Jan 29.

Abstract

We employ the CRISPR-Cas system of Streptococcus pyogenes as programmable RNA-guided endonucleases (RGENs) to cleave DNA in a targeted manner for genome editing in human cells. We show that complexes of the Cas9 protein and artificial chimeric RNAs efficiently cleave two genomic sites and induce indels with frequencies of up to 33%.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism
  • Base Sequence
  • CRISPR-Cas Systems
  • DNA Cleavage
  • DNA, Recombinant / genetics
  • DNA, Recombinant / metabolism
  • Endonucleases / genetics*
  • Endonucleases / metabolism
  • Genes, Reporter
  • Genetic Engineering / methods*
  • Genome, Human*
  • Histocompatibility Antigens / genetics
  • Histocompatibility Antigens / metabolism
  • Humans
  • Luminescent Proteins / genetics
  • Luminescent Proteins / metabolism
  • Molecular Sequence Data
  • Mutagenesis, Site-Directed
  • RNA, Guide / genetics*
  • RNA, Guide / metabolism
  • Receptors, CCR5 / genetics
  • Receptors, CCR5 / metabolism
  • Streptococcus pyogenes / enzymology
  • Streptococcus pyogenes / genetics
  • Zinc Fingers

Substances

  • Bacterial Proteins
  • C4BPB protein, human
  • DNA, Recombinant
  • Histocompatibility Antigens
  • Luminescent Proteins
  • RNA, Guide
  • Receptors, CCR5
  • Endonucleases