Site-specific cleavage of a yeast chromosome by oligonucleotide-directed triple-helix formation.
Site-specific cleavage of a yeast chromosome by oligonucleotide-directed triple-helix formation.
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通过寡核苷酸引导的三螺旋形成对酵母染色体进行位点特异性切割。
DOI：
10.1126/science.2195655
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发表时间：
1990
期刊：
影响因子：
--
通讯作者：
Dervan,PB
中科院分区：
文献类型：
--
作者：
Strobel,SA;Dervan,PB
配备有EDTA螯合铁的寡核苷酸可以通过三螺旋形成特异性结合双链DNA，并在大于12个碱基对的结合位点产生双链切割。为了证明三螺旋形成是一种可行的化学方法，用于大基因组DNA的位点特异性切割，在5′和3′末端具有EDTA双金属Fe的寡核苷酸靶向于酿酒酵母染色体III的340个组氨酸酶对中的20个碱基对序列。观察到正确大小和位置的双链切割产物，表明寡核苷酸结合并切割了近14兆碱基对DNA中的靶位点。由于三螺旋结构的形成有可能成为DNA识别的通用解决方案，因此这一结果对染色体的物理作图具有重要意义。
Oligonucleotides equipped with EDTA⋅Fe can bind specifically to duplex DNA by triple-helix formation and produce double-strand cleavage at binding sites greater than 12 base pairs in size. To demonstrate that oligonucleotide-directed triple-helix formation is a viable chemical approach for the site-specific cleavage of large genomic DNA, an oligonucleotide with EDTA⋅Fe at the 5′ and 3′ ends was targeted to a 20-base pair sequence in the 340-kilobase pair chromosome III ofSaccharomyces cerevisiae. Double-strand cleavage products of the correct size and location were observed, indicating that the oligonucleotide bound and cleaved the target site among almost 14 megabase pairs of DNA. Because oligonucleotide-directed triple-helix formation has the potential to be a general solution for DNA recognition, this result has implications for physical mapping of chromosomes.