New strategy for the synthesis of chemically modified RNA constructs exemplified by hairpin and hammerhead ribozymes
New strategy for the synthesis of chemically modified RNA constructs exemplified by hairpin and hammerhead ribozymes
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DOI：
10.1073/pnas.1006447107
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发表时间：
2010-08-31
影响因子：
11.1
通讯作者：
Brown, Tom
中科院分区：
文献类型：
--
作者：
El-Sagheer, Afaf H.;Brown, Tom
CuAAC反应（点击化学）已与固相合成结合使用，以产生长度约为100个核苷酸的催化活性发夹核酶。通过相邻核碱基的跨链连接成功地以高效率共价连接预合成的RNA链（反式连接）。在另一种策略中，采用链内点击连接来产生在催化位点处含有新型核酸骨架模拟物的功能性锤头状核酶（顺式连接）。通过固相合成和点击连接的组合合成长RNA链的能力是RNA化学的重要补充。它与大量的位点特异性修饰相容，并且适用于许多生物学上重要的RNA分子的合成。
The CuAAC reaction (click chemistry) has been used in conjunction with solid-phase synthesis to produce catalytically active hairpin ribozymes around 100 nucleotides in length. Cross-strand ligation through neighboring nucleobases was successful in covalently linking presynthesized RNA strands with high efficiency (trans-ligation). In an alternative strategy, intrastrand click ligation was employed to produce a functional hammerhead ribozyme containing a novel nucleic acid backbone mimic at the catalytic site (cis-ligation). The ability to synthesize long RNA strands by a combination of solid-phase synthesis and click ligation is an important addition to RNA chemistry. It is compatible with a plethora of site-specific modifications and is applicable to the synthesis of many biologically important RNA molecules.