Variety of nucleotide polymerase mutants aiming to synthesize modified RNA
Variety of nucleotide polymerase mutants aiming to synthesize modified RNA
复制标题
旨在合成修饰RNA的多种核苷酸聚合酶突变体
DOI：
10.1002/cbic.202100004
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发表时间：
2021
期刊：
影响因子：
3.2
通讯作者：
Hiroshi Abe
中科院分区：
文献类型：
--
作者：
Sana Ohashi;Fumitaka Hashiya;Hiroshi Abe
已经进行了大量的努力来开发利用合成RNA捕获靶分子的治疗性RNA适体。然而，确保RNA适体对内在核酸酶具有抗性仍然是一个问题，并限制了它们作为治疗剂的用途。对RNA的2′糖部分进行化学修饰可以有效地提高其稳定性，并且可以通过使用修饰的亚磷酰胺进行化学合成来实现;然而，这种方法不适合制备长RNA分子。虽然重组核苷酸聚合酶可以转录RNA，但这些聚合酶不能合成修饰的RNA，因为它们不识别2′修饰的核苷三磷酸。本文综述了几种能耐受2′-糖基修饰的底物合成RNA的聚合酶突变体，以及利用体外转录技术制备高效率的化学修饰RNA所需克服的问题。
Significant efforts have been made to develop therapeutic RNA aptamers that exploit synthetic RNA to capture target molecules. However, ensuring RNA aptamers are resistant against intrinsic nucleases remains an issue and restricts their use as therapeutics. Introduction of chemical modifications to the 2′ sugar moiety of RNA improves their stability effectively and can be achieved by chemical synthesis using modified phosphoramidites; however, this approach is not suitable for preparing long RNA molecules. Although recombinant nucleotide polymerases can transcribe RNA, these polymerases cannot synthesize modified RNA because they do not recognize 2′ modified nucleoside triphosphates. In this review, we focus on several polymerase mutants that tolerate substrates containing modifications of the 2′ sugar moiety to synthesize RNA, and the problems that must be overcome to prepare chemically modified RNA with high efficacy byin vitrotranscription.