Molecular and Cellular Biology Program, University of Washington, Seattle, Washington, USA.
Nat Methods. 2012 Oct;9(10):973-5. doi: 10.1038/nmeth.2177. Epub 2012 Sep 2.
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.
靶向 DNA 双链断裂由罕见切割的设计内切酶引入,可以通过参与诱变非同源末端连接 DNA 修复途径来用于基因敲除应用。然而,内切酶介导的 DNA 断裂通常受到精确修复的限制,从而限制了靶向基因组编辑的效率。为了解决这个问题,我们将设计内切酶与 DNA 末端处理酶偶联,以驱动诱变断裂的解决,从而使基因敲除率提高了 25 倍。