Coupling endonucleases with DNA end-processing enzymes to drive gene disruption

Nat Methods. 2012 Oct;9(10):973-5. doi: 10.1038/nmeth.2177. Epub 2012 Sep 2.

Abstract

Targeted DNA double-strand breaks introduced by rare-cleaving designer endonucleases can be harnessed for gene disruption applications by engaging mutagenic nonhomologous end-joining DNA repair pathways. However, endonuclease-mediated DNA breaks are often subject to precise repair, which limits the efficiency of targeted genome editing. To address this issue, we coupled designer endonucleases to DNA end-processing enzymes to drive mutagenic break resolution, achieving up to 25-fold enhancements in gene disruption rates.

Publication types

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

MeSH terms

  • Animals
  • DNA Breaks, Double-Stranded*
  • DNA End-Joining Repair
  • DNA Repair
  • Endonucleases / physiology*
  • Exodeoxyribonucleases / physiology
  • HEK293 Cells
  • Humans
  • Mice
  • Phosphoproteins / physiology
  • Receptors, CCR5 / physiology

Substances

  • Phosphoproteins
  • Receptors, CCR5
  • Endonucleases
  • Exodeoxyribonucleases
  • TREX2 protein, human