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RS-1 Enhances CRISPR/Cas9- And TALEN-mediated Knock-In Efficiency

RS-1 Enhances CRISPR/Cas9- And TALEN-mediated Knock-In Efficiency

Jun Song, Dongshan Yang, Jie Xu, Tianqing Zhu, Y Eugene Chen, Jifeng Zhang

Nat Commun. 2016 Jan 28;7:10548.

PMID: 26817820

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.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
AP312756744	RS-1		312756-74-4	Price

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