Novel RNA synthesis method using 5′-O-silyl-2′-O-orthoester protecting groups
Novel RNA synthesis method using 5′-O-silyl-2′-O-orthoester protecting groups
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DOI：
10.1021/ja980730v
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发表时间：
1998-11-18
影响因子：
15
通讯作者：
Caruthers, MH
中科院分区：
文献类型：
--
作者：
Scaringe, SA;Wincott, FE;Caruthers, MH
随着研究揭示了RNA的多种生物学功能，常规合成RNA的能力变得越来越重要。在过去的25年里，人们探索了许多合成RNA的化学策略。大多数方法都集中在保留5′-O-二甲氧基三苯甲基（DMT）醚并添加相容的2′-羟基保护基，如氟化物不稳定的甲硅烷基醚、2个光不稳定部分、3或酸不稳定的缩醛。4缩醛类化合物具有许多吸引人的特性，但在相同的合成策略中成功地利用2′-O-缩醛和5′-O-DMT醚需要一个微妙的平衡。5因此，其他方法包括保留2′-O-缩醛，同时取代5′-O-DMT。6若干审查进一步记录了这些战略。7在迄今为止报道的所有RNA合成方法中，5′-O-DMT-2′-O-叔丁基二甲基甲硅烷基（TBDMS）和5′-O-DMT-2′-O-[1-（2-氟苯基）-4-甲氧基哌啶-4-基]（FPMP）化学是商业上提供的。不幸的是，这两种方法都不允许RNA合成像DNA那样常规和可靠。目前的方法能够以可接受的产率和质量合成RNA，但似乎需要高水平的技能来提供足够的结果。需要和愿望存在更强大的RNA合成方法，始终产生更高质量的RNA. While大多数以前的方法是DNA方法的改编，我们专注于一个de noVo策略，并询问什么是最佳的RNA。根据文献，最终2′-O-脱保护的最理想条件是弱酸性水溶液条件。使用轻度酸不稳定的2′-O-基团的障碍是5′-O-DMT基团，它在类似条件下被除去。我们的研究成功地开发了用于保护5′-羟基的甲硅烷基醚。8这些保护基可以用氟化物去除
The ability to routinely synthesize RNA has become increasingly important as research reveals the multitude of RNA’s biological functions. 1 Over the past 25 years, many chemical strategies have been explored for synthesizing RNA. Most approaches have focused on retaining the 5′-O-dimethoxytrityl (DMT) ether and adding a compatible 2′-hydroxyl protecting group such as fluoride-labile silyl ethers, 2 photolabile moieties, 3 or acid-labile acetals. 4 The acetals have exhibited many attractive features, but a delicate balance has been required to successfully utilize the 2′-O-acetals and the 5′-O-DMT ether in the same synthesis strategy. 5 Hence, other approaches have involved retaining the 2′-O-acetal while replacing the 5′-O-DMT. 6 Several reviews further document these strategies. 7 Of all of the RNA synthesis methods reported to date, the 5′-O-DMT-2′-O-tert-butyldimethylsilyl (TBDMS) and the 5′-O-DMT-2′-O-[1-(2-fluorophenyl)-4-methoxypiperidin-4-yl](FPMP) chemistries are offered commercially. Unfortunately, neither allows RNA synthesis to be as routine and dependable as DNA. The current methods enable the synthesis of RNA in acceptable yields and quality, but a high level of skill appears to be required to deliver adequate results. The need and desire exists for more robust RNA synthesis methods which consistently produce higher quality RNA.Whereas most previous approaches were adaptations of DNA methodologies, we focused on a de noVo strategy and asked what would be optimal for RNA. According to the literature, the most desirable conditions for the final 2′-O-deprotection would be mildly acidic aqueous conditions. The obstacle to using mildly acid-labile 2′-O-groups has been the 5′-O-DMT group, which is removed under similar conditions. Our investigations led to the successful development of silyl ethers for protection of the 5′-hydroxyl. 8 These protecting groups can be removed with fluoride