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

Author

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  • Jun Song

    (Center for Advanced Models for Translational Sciences and Therapeutics, University of Michigan Medical Center)

  • Dongshan Yang

    (Center for Advanced Models for Translational Sciences and Therapeutics, University of Michigan Medical Center)

  • Jie Xu

    (Center for Advanced Models for Translational Sciences and Therapeutics, University of Michigan Medical Center)

  • Tianqing Zhu

    (Center for Advanced Models for Translational Sciences and Therapeutics, University of Michigan Medical Center)

  • Y. Eugene Chen

    (Center for Advanced Models for Translational Sciences and Therapeutics, University of Michigan Medical Center)

  • Jifeng Zhang

    (Center for Advanced Models for Translational Sciences and Therapeutics, University of Michigan Medical Center)

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.

Suggested Citation

  • Jun Song & Dongshan Yang & Jie Xu & Tianqing Zhu & Y. Eugene Chen & Jifeng Zhang, 2016. "RS-1 enhances CRISPR/Cas9- and TALEN-mediated knock-in efficiency," Nature Communications, Nature, vol. 7(1), pages 1-7, April.
    • Handle: RePEc:nat:natcom:v:7:y:2016:i:1:d:10.1038_ncomms10548
    DOI: 10.1038/ncomms10548
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    Cited by:

    1. Lukas Möller & Eric J. Aird & Markus S. Schröder & Lena Kobel & Lucas Kissling & Lilly van de Venn & Jacob E. Corn, 2022. "Recursive Editing improves homology-directed repair through retargeting of undesired outcomes," Nature Communications, Nature, vol. 13(1), pages 1-10, December.
    2. Michael Kosicki & Felicity Allen & Frances Steward & Kärt Tomberg & Yangyang Pan & Allan Bradley, 2022. "Cas9-induced large deletions and small indels are controlled in a convergent fashion," Nature Communications, Nature, vol. 13(1), pages 1-11, December.

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