Resetting the Site: Redirecting Integration of an Insertion Sequence in a Predictable Way
Resetting the Site: Redirecting Integration of an Insertion Sequence in a Predictable Way
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DOI：
10.1016/j.molcel.2009.05.017
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发表时间：
2009-06-12
期刊：
影响因子：
16
通讯作者：
Chandler, Michael
中科院分区：
文献类型：
--
作者：
Guynet, Catherine;Achard, Adeline;Chandler, Michael
尽管目标位点选择在合理的转座子介导的基因传递中很重要，但它是 DNA 转座的一个复杂且知之甚少的方面。尽管大多数转座子基本上随机地选择靶位点，或者具有一些轻微的序列或结构偏好，但来自幽门螺杆菌的插入序列 IS608（使用单链 DNA 进行转座）始终仅插入 TTAC 四核苷酸的 3'。我们对 IS608 转座机制的研究结果表明，转座酶通过选择转座子 DNA 的内部片段来识别其目标位点，并利用它通过碱基配对来特异性识别目标位点。这提出了一种将 IS608 转座重定向到新目标位点的方法。正如我们在这里所演示的，我们现在可以以可预测的方式在体外和体内直接插入各种不同的选定靶序列。
Target site choice is a complex and poorly understood aspect of DNA transposition despite its importance in rational transposon-mediated gene delivery. Though most transposons choose target sites essentially randomly or with some slight sequence or structural preferences, insertion sequence IS608 from Helicobacter pylori, which transposes using single-stranded DNA, always inserts just 3' of a TTAC tetranucleotide. Our results from studies on the IS608 transposition mechanism demonstrated that the transposase recognizes its target site by co-opting an internal segment of transposon DNA and utilizes it for specific recognition of the target sites through base-pairing. This suggested a way to redirect IS608 transposition to novel target sites. As we demonstrate here, we can now direct insertions in a predictable way into a variety of different chosen target sequences, both in vitro and in vivo.