Front Cover: A T7 RNA Polymerase Mutant Enhances the Yield of 5′‐Thienoguanosine‐Initiated RNAs (ChemBioChem 2/2018),ChemBioChem

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Front Cover: A T7 RNA Polymerase Mutant Enhances the Yield of 5′‐Thienoguanosine‐Initiated RNAs (ChemBioChem 2/2018)
ChemBioChem ( IF 2.6 ) Pub Date : 2018-01-09 , DOI: 10.1002/cbic.201700676
Seth Lyon ₁ , Venkat Gopalan ₁

Affiliation

The front cover picture shows a one‐pot multi‐enzyme approach that was developed to enhance the yield and purity of RNAs incorporating 5′‐thienoguanosine (^thG). In vitro transcription has been employed to introduce chemical handles and modifications into RNAs, but is typically constrained by poor yields of the modified RNAs. A two‐pronged solution to this long‐standing problem is depicted here: 1) the use of a P266L mutant of T7 RNA polymerase (PDB ID: 3E2E) decreases abortive transcription and improves the total RNA yield as well as percentage incorporation of ^thG (a fluorescent guanosine surrogate), and 2) the addition of 5′‐pyrophosphohydrolase (RppH) and exoribonuclease I (Xrn1) degrades GTP‐initiated RNAs. This approach results in a better yield of near‐homogenous 5′‐^thG‐incorporated RNA, and should have broad utility for the synthesis of 5′‐modified RNAs. More information can be found in the communication by S. Lyon and V. Gopalan on page 142 in Issue 2, 2018 (DOI: 10.1002/cbic.201700538).

中文翻译：

封面：T7 RNA聚合酶突变体可提高5'-硫鸟嘌呤引发的RNA的产量（ChemBioChem 2/2018）

封面图片显示了一种单罐多酶方法，该方法旨在提高掺入5'-硫代鸟苷（^th G）的RNA的产量和纯度。体外转录已被用于将化学处理和修饰引入RNA中，但通常受到修饰RNA产量低的限制。甲双叉解决这个长期存在的问题是这里所示：1）使用T7 RNA聚合酶的突变体P266L的（PDB ID：3E2E）减小无效转录，并提高了总RNA产量以及百分比掺入^第G（一种荧光鸟嘌呤替代物），以及2）添加5'-焦磷酸水解酶（RppH）和核糖核酸外切酶I（Xrn1）可以降解GTP起始的RNA。这种做法的结果的更好的产率近乎均匀的5'-^次G-掺入RNA，并应具有用于5'-修饰RNA的合成具有广泛的用途。可以在2018年第2期第142页的S.Lyon和V.Gopalan的通信中找到更多信息（DOI：10.1002 / cbic.201700538）。

更新日期：2018-01-09

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