Skip to main page content
Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2015 May;33(5):543-8.
doi: 10.1038/nbt.3198. Epub 2015 Mar 24.

Increasing the Efficiency of Homology-Directed Repair for CRISPR-Cas9-induced Precise Gene Editing in Mammalian Cells

Affiliations

Increasing the Efficiency of Homology-Directed Repair for CRISPR-Cas9-induced Precise Gene Editing in Mammalian Cells

Van Trung Chu et al. Nat Biotechnol. .

Erratum in

Abstract

The insertion of precise genetic modifications by genome editing tools such as CRISPR-Cas9 is limited by the relatively low efficiency of homology-directed repair (HDR) compared with the higher efficiency of the nonhomologous end-joining (NHEJ) pathway. To enhance HDR, enabling the insertion of precise genetic modifications, we suppressed the NHEJ key molecules KU70, KU80 or DNA ligase IV by gene silencing, the ligase IV inhibitor SCR7 or the coexpression of adenovirus 4 E1B55K and E4orf6 proteins in a 'traffic light' and other reporter systems. Suppression of KU70 and DNA ligase IV promotes the efficiency of HDR 4-5-fold. When co-expressed with the Cas9 system, E1B55K and E4orf6 improved the efficiency of HDR up to eightfold and essentially abolished NHEJ activity in both human and mouse cell lines. Our findings provide useful tools to improve the frequency of precise gene modifications in mammalian cells.

Similar articles

See all similar articles

Cited by 328 articles

See all "Cited by" articles

References

    1. Cell. 2005 Dec 16;123(6):1013-23 - PubMed
    1. J Virol. 2011 Mar;85(5):2201-11 - PubMed
    1. Aging (Albany NY). 2011 Apr;3(4):395-406 - PubMed
    1. Cell. 2012 Dec 21;151(7):1474-87 - PubMed
    1. Cell. 2009 Oct 30;139(3):573-86 - PubMed

Publication types

LinkOut - more resources

Feedback