Specific mutations induced by triplex-forming oligonucleotides in mice
Specific mutations induced by triplex-forming oligonucleotides in mice
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DOI：
10.1126/science.290.5491.530
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发表时间：
2000-10-20
期刊：
影响因子：
56.9
通讯作者：
Glazer, PM
中科院分区：
文献类型：
--
作者：
Vasquez, KM;Narayanan, L;Glazer, PM
三聚体形成寡核苷酸(TFOS)识别并结合特定的双链DNA序列，已被广泛用于修饰细胞中的基因功能。虽然生殖系突变可以通过胚胎干细胞技术进行整合，但在将突变引入活生物的体细胞方面进展甚微。在这里，我们证明了TFOS可以在成年小鼠的体细胞中诱导特定基因组位置的突变。通过使用携带supF和CLL报告基因的染色体副本的转基因小鼠，突变检测变得容易。接受supF靶向TFO治疗的小鼠的supF基因突变频率比接受扰乱序列控制寡聚体治疗的小鼠高约五倍，在使用这两种寡核苷酸的对照基因(CLL)中都没有检测到突变。这些结果表明，可以在完好的动物身上完成特定部位的、TFO导向的基因组修饰。
Tripler-forming oligonucleotides (TFOs) recognize and bind to specific duplex DNA sequences and have been used extensively to modify gene function in cells. Although germ line mutations can be incorporated by means of embryonic stem cell technology, little progress has been made toward introducing mutations in somatic cells of living organisms. Here we demonstrate that TFOs can induce mutations at specific genomic sites in somatic cells of adult mice. Mutation detection was facilitated by the use of transgenic mice bearing chromosomal copies of the supF and cll reporter genes. Mice treated with a supF-targeted TFO displayed about fivefold greater mutation frequencies in the supF gene compared with mice treated with a scrambled sequence control oligomer, No mutagenesis was detected in the control gene (cll) with either oligonucleotide. These results demonstrate that site-specific, TFO-directed genome modification can be accomplished in intact animals.