Department of Medicine, University of Washington, Seattle, WA, 98195, USA and Department of Biochemistry, University of Washington, Seattle, WA, 98195, USA.
Nucleic Acids Res. 2014 Mar;42(5):3119-24. doi: 10.1093/nar/gkt1303. Epub 2013 Dec 25.
DNA mismatches that occur between vector homology arms and chromosomal target sequences reduce gene targeting frequencies in several species; however, this has not been reported in human cells. Here we demonstrate that even a single mismatched base pair can significantly decrease human gene targeting frequencies. In addition, we show that homology arm polymorphisms can be used to direct allele-specific targeting or to improve unfavorable vector designs that introduce deletions.
在几种物种中，载体同源臂和染色体靶序列之间发生的 DNA 错配会降低基因靶向频率；然而，在人类细胞中尚未有报道。在这里，我们证明了即使单个错配碱基对也可以显著降低人类基因靶向频率。此外，我们还表明，同源臂多态性可用于指导等位基因特异性靶向，或改进引入缺失的不利载体设计。
