Coupling endonucleases with DNA end-processing enzymes to drive gene disruption.

Certo, Michael T; Gwiazda, Kamila S; Kuhar, Ryan; Sather, Blythe; Curinga, Gabrielle; Mandt, Tyler; Brault, Michelle; Lambert, Abigail R et al. · Nat Methods · 2012

basic_science · Level V

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Abstract

Targeted DNA double-strand breaks introduced by rare-cleaving designer endonucleases can be harnessed for gene disruption applications by engaging mutagenic nonhomologous end-joining DNA repair pathways. However, endonuclease-mediated DNA breaks are often subject to precise repair, which limits the efficiency of targeted genome editing. To address this issue, we coupled designer endonucleases to DNA end-processing enzymes to drive mutagenic break resolution, achieving up to 25-fold enhancements in gene disruption rates.

Medical subject headings