Simultaneous targeted exchange of two nucleotides by single-stranded oligonucleotides clusters within a region of about fourteen nucleotides
Simultaneous targeted exchange of two nucleotides by single-stranded oligonucleotides clusters within a region of about fourteen nucleotides
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DOI：
10.1186/1471-2199-9-14
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发表时间：
2008-01-28
影响因子：
--
通讯作者：
Kaufmann, Dieter
中科院分区：
文献类型：
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作者：
Hegele, Heike;Wuepping, Matthias;Kaufmann, Dieter
背景：用基因特异性的单链寡核苷酸转染细胞可以诱导靶基因中一个或两个核苷酸的靶向交换。为了描述所涉及的dna修复机制的特征，我们检查了单链寡核苷酸同时交换两个核苷酸的最大距离。所选择的实验系统是纠正仓鼠细胞系中的hrt点突变，在距离第一个交换位置可变距离处产生额外的核苷酸交换，并调查同时核苷酸交换的速率。结果：两个交换位置之间的距离越小，同时交换的概率越高。同时检测到的核苷酸交换被发现聚集在第一个交换位置的上游和下游约14个核苷酸的区域。结论：我们认为参与修复目标DNA链的机制仅利用单链寡核苷酸的短序列，它可能被物理地结合到DNA中或用作修复过程的基质。
Background: Transfection of cells with gene-specific, single-stranded oligonucleotides can induce the targeted exchange of one or two nucleotides in the targeted gene. To characterize the features of the DNA-repair mechanisms involved, we examined the maximal distance for the simultaneous exchange of two nucleotides by a single-stranded oligonucleotide. The chosen experimental system was the correction of a hprt-point mutation in a hamster cell line, the generation of an additional nucleotide exchange at a variable distance from the first exchange position and the investigation of the rate of simultaneous nucleotide exchanges.Results: The smaller the distance between the two exchange positions, the higher was the probability of a simultaneous exchange. The detected simultaneous nucleotide exchanges were found to cluster in a region of about fourteen nucleotides upstream and downstream from the first exchange position.Conclusion: We suggest that the mechanism involved in the repair of the targeted DNA strand utilizes only a short sequence of the single-stranded oligonucleotide, which may be physically incorporated into the DNA or be used as a matrix for a repair process.