1. Academic Validation
  2. RS-1 enhances CRISPR/Cas9- and TALEN-mediated knock-in efficiency

RS-1 enhances CRISPR/Cas9- and TALEN-mediated knock-in efficiency

  • Nat Commun. 2016 Jan 28;7:10548. doi: 10.1038/ncomms10548.
Jun Song 1 Dongshan Yang 1 Jie Xu 1 Tianqing Zhu 1 Y Eugene Chen 1 Jifeng Zhang 1
Affiliations

Affiliation

  • 1 Center for Advanced Models for Translational Sciences and Therapeutics, University of Michigan Medical Center, Ann Arbor, Michigan 48109, USA.
Abstract

Zinc-finger nuclease, transcription activator-like effector nuclease and CRISPR (clustered regularly interspaced short palindromic repeats)/Cas9 (CRISPR-associated protein 9) are becoming major tools for genome editing. Importantly, knock-in in several non-rodent species has been finally achieved thanks to these customizable nucleases; yet the rates remain to be further improved. We hypothesize that inhibiting non-homologous end joining (NHEJ) or enhancing homology-directed repair (HDR) will improve the nuclease-mediated knock-in efficiency. Here we show that the in vitro application of an HDR enhancer, RS-1, increases the knock-in efficiency by two- to five-fold at different loci, whereas NHEJ inhibitor SCR7 has minimal effects. We then apply RS-1 for animal production and have achieved multifold improvement on the knock-in rates as well. Our work presents tools to nuclease-mediated knock-in animal production, and sheds LIGHT on improving gene-targeting efficiencies on pluripotent stem cells.

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