哺乳动物细胞中三链螺旋靶向DNA损伤诱导的重组。
Recombination induced by triple-helix-targeted DNA damage in mammalian cells.
作者信息
Faruqi A F, Seidman M M, Segal D J, Carroll D, Glazer P M
机构信息
Department of Therapeutic Radiology, Yale University School of Medicine, New Haven, Connecticut 06520-8040, USA.
出版信息
Mol Cell Biol. 1996 Dec;16(12):6820-8. doi: 10.1128/MCB.16.12.6820.
Abstract
Gene therapy has been hindered by the low frequency of homologous recombination in mammalian cells. To stimulate recombination, we investigated the use of triple-helix-forming oligonucleotides (TFOs) to target DNA damage to a selected site within cells. By treating cells with TFOs linked to psoralen, recombination was induced within a simian virus 40 vector carrying two mutant copies of the supF tRNA reporter gene. Gene conversion events, as well as mutations at the target site, were also observed. The variety of products suggests that multiple cellular pathways can act on the targeted damage, and data showing that the triple helix can influence these pathways are presented. The ability to specifically induce recombination or gene conversion within mammalian cells by using TFOs may provide a new research tool and may eventually lead to novel applications in gene therapy.
摘要
基因治疗一直受到哺乳动物细胞中同源重组频率低的阻碍。为了刺激重组，我们研究了使用三链形成寡核苷酸（TFO）将DNA损伤靶向细胞内选定位点的方法。通过用与补骨脂素相连的TFO处理细胞，在携带supF tRNA报告基因两个突变拷贝的猿猴病毒40载体中诱导了重组。还观察到了基因转换事件以及靶位点的突变。多种产物表明多种细胞途径可作用于靶向损伤，并展示了表明三链螺旋可影响这些途径的数据。利用TFO在哺乳动物细胞中特异性诱导重组或基因转换的能力可能提供一种新的研究工具，并最终可能导致基因治疗中的新应用。
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