Padlock oligonucleotides for duplex DNA based on sequence-specific triple helix formation
Padlock oligonucleotides for duplex DNA based on sequence-specific triple helix formation
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DOI：
10.1073/pnas.96.19.10603
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发表时间：
1999-09-14
影响因子：
11.1
通讯作者：
Hélène, C
中科院分区：
文献类型：
--
作者：
Escudé, C;Garestier, T;Hélène, C
由于三螺旋的形成，寡核苷酸在双链DNA周围循环。已知的是，短的寡核苷酸能够通过与DNA主槽结合形成DNA三螺旋。在通过三螺旋形成对双链DNA靶标进行序列特异性识别后，三倍体形成的寡核苷酸的末端通过T4 DNA连接酶的作用连接在一起，从而产生与含有靶标序列的质粒相连的环状DNA分子。双链DNA序列的标记在没有对该序列进行任何化学或酶修饰的情况下进行。这些“挂锁”寡核苷酸提供了一种工具，可以以不可逆的方式将非共价标签连接到超螺旋质粒或其他双链DNA上。这种复合体可能会在开发双链DNA检测的新技术或用于基因治疗的质粒递送方法中得到应用。
An oligonucleotide was circularized around double-stranded DNA thanks to triple helix formation. Short oligonucleotides are known to be able to form DNA triple helices by binding into the DNA major groove at an oligopurine oligopyrimidine sequence. After sequence-specific recognition of a double-stranded DNA target through triple helix formation, the ends of the tripler-forming oligonucleotide were joined through the action of T4 DNA ligase, thus creating a circular DNA molecule catenated to the plasmid containing the target sequence. The labeling of the double-stranded DNA sequence has been carried out without any chemical or enzymatic modification of this sequence. These "padlock" oligonucleotides provide a tool to attach a noncovalent tag in an irreversible way to supercoiled plasmid or other double-stranded DNAs. Such a complex may find applications in the development of new techniques for duplex DNA detection or plasmid delivery methods for gene therapy.