Increasing the efficiency of precise genome editing with CRISPR-Cas9 by inhibition of nonhomologous end joining View Full Text


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

2015-05

AUTHORS

Takeshi Maruyama, Stephanie K Dougan, Matthias C Truttmann, Angelina M Bilate, Jessica R Ingram, Hidde L Ploegh

ABSTRACT

Methods to introduce targeted double-strand breaks (DSBs) into DNA enable precise genome editing by increasing the rate at which externally supplied DNA fragments are incorporated into the genome through homologous recombination. The efficiency of these methods is limited by nonhomologous end joining (NHEJ), an alternative DNA repair pathway that competes with homology-directed repair (HDR). To promote HDR at the expense of NHEJ, we targeted DNA ligase IV, a key enzyme in the NHEJ pathway, using the inhibitor Scr7. Scr7 treatment increased the efficiency of HDR-mediated genome editing, using Cas9 in mammalian cell lines and in mice for all four genes examined, up to 19-fold. This approach should be applicable to other customizable endonucleases, such as zinc finger nucleases and transcription activator-like effector nucleases, and to nonmammalian cells with sufficiently conserved mechanisms of NHEJ and HDR. More... »

PAGES

538-542

Identifiers

URI

http://scigraph.springernature.com/pub.10.1038/nbt.3190

DOI

http://dx.doi.org/10.1038/nbt.3190

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1017538548

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/25798939


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