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


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

2016-01-28

AUTHORS

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

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. More... »

PAGES

10548

Identifiers

URI

http://scigraph.springernature.com/pub.10.1038/ncomms10548

DOI

http://dx.doi.org/10.1038/ncomms10548

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1042173867

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/26817820


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