Influence of DNA delivery method on gene targeting frequencies in human cells.

R J Yáñez, A C Porter
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引用次数: 35

Abstract

Gene targeting can be used for genetic studies of human cell lines and has significant potential for somatic cell gene therapy. These applications are however restricted by the low frequency of homologous recombination in higher eukaryotes compared to the relatively efficient nonhomologous integration of transfected DNA into the genome. As part of our attempts to overcome this problem, we compared two widely used transfection methods for their efficiency in gene targeting. To our surprise we found that, for conditions that render similar frequencies of nonhomologous integrants, lipofection is much less efficient than electroporation in generating targeted clones. This suggests that nonhomologous and homologous recombination have different requirements for DNA delivery in human cells.

DNA传递方式对人细胞基因靶向频率的影响。
基因靶向可以用于人类细胞系的遗传研究,在体细胞基因治疗方面具有重要的潜力。然而,与转染DNA相对高效的非同源整合到基因组中相比,这些应用受到高等真核生物中同源重组频率较低的限制。作为我们尝试克服这个问题的一部分,我们比较了两种广泛使用的转染方法在基因靶向方面的效率。令我们惊讶的是,我们发现,在非同源整合物频率相似的条件下,脂肪转染在产生目标克隆方面的效率远低于电穿孔。这表明非同源重组和同源重组对人类细胞DNA传递有不同的要求。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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