Access to capped RNAs by chemical ligation†

IF 4.2 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY RSC Chemical Biology Pub Date : 2024-09-13 DOI:10.1039/D4CB00165F
Karolina Bartosik and Ronald Micura
{"title":"Access to capped RNAs by chemical ligation†","authors":"Karolina Bartosik and Ronald Micura","doi":"10.1039/D4CB00165F","DOIUrl":null,"url":null,"abstract":"<p >A distinctive feature of eukaryotic mRNAs is the presence of a cap structure at the 5′ end. The typical cap consists of 7-methylguanosine linked to the first transcribed nucleotide through a 5′,5′-triphosphate bridge. It plays a key role in many processes in eukaryotic cells, including splicing, intracellular transport, initiation of translation and turnover. Synthetic capped oligonucleotides have served as useful tools for elucidating these physiological processes. In addition, cap mimics with artificial modifications are of interest for the design of mRNA-based therapeutics and vaccines. While the short cap mimics can be obtained by chemical synthesis, the preparation of capped analogs of mRNA length is still challenging and requires templated enzymatic ligation of synthetic RNA fragments. To increase the availability of capped mRNA analogs, we present here a practical and non-templated approach based on the use of click ligation resulting in RNAs bearing a single triazole linkage within the oligo-phosphate backbone. Capped RNA fragments with up to 81 nucleotides in length have thus been obtained in nanomolar yields and are in demand for biochemical, spectroscopic or structural studies.</p>","PeriodicalId":40691,"journal":{"name":"RSC Chemical Biology","volume":null,"pages":null},"PeriodicalIF":4.2000,"publicationDate":"2024-09-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.rsc.org/en/content/articlepdf/2024/cb/d4cb00165f?page=search","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"RSC Chemical Biology","FirstCategoryId":"1085","ListUrlMain":"https://pubs.rsc.org/en/content/articlelanding/2024/cb/d4cb00165f","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"BIOCHEMISTRY & MOLECULAR BIOLOGY","Score":null,"Total":0}
引用次数: 0

Abstract

A distinctive feature of eukaryotic mRNAs is the presence of a cap structure at the 5′ end. The typical cap consists of 7-methylguanosine linked to the first transcribed nucleotide through a 5′,5′-triphosphate bridge. It plays a key role in many processes in eukaryotic cells, including splicing, intracellular transport, initiation of translation and turnover. Synthetic capped oligonucleotides have served as useful tools for elucidating these physiological processes. In addition, cap mimics with artificial modifications are of interest for the design of mRNA-based therapeutics and vaccines. While the short cap mimics can be obtained by chemical synthesis, the preparation of capped analogs of mRNA length is still challenging and requires templated enzymatic ligation of synthetic RNA fragments. To increase the availability of capped mRNA analogs, we present here a practical and non-templated approach based on the use of click ligation resulting in RNAs bearing a single triazole linkage within the oligo-phosphate backbone. Capped RNA fragments with up to 81 nucleotides in length have thus been obtained in nanomolar yields and are in demand for biochemical, spectroscopic or structural studies.

Abstract Image

Abstract Image

查看原文
分享 分享
微信好友 朋友圈 QQ好友 复制链接
本刊更多论文
通过化学连接获取封端 RNA
真核生物 mRNA 的一个显著特点是其 5′末端有一个帽子结构。典型的帽结构由 7-甲基鸟苷酸组成,通过 5′、5′-三磷酸桥与第一个转录核苷酸相连。它在真核细胞的许多过程中都起着关键作用,包括剪接、细胞内运输、启动翻译和转换。合成的带帽寡核苷酸是阐明这些生理过程的有用工具。此外,具有人工修饰的帽子模拟物对设计基于 mRNA 的疗法和疫苗也很有意义。虽然可以通过化学合成获得短的帽状模拟物,但制备 mRNA 长度的帽状类似物仍然具有挑战性,需要对合成的 RNA 片段进行模板化的酶连接。为了提高带帽 mRNA 类似物的可用性,我们在此介绍一种实用的非模板方法,该方法基于点击连接,从而在寡聚磷酸骨架内产生带有单个三唑连接的 RNA。因此,我们以纳摩尔产量获得了长度达 81 个核苷酸的带帽 RNA 片段,这些片段是生化、光谱或结构研究的必需品。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
求助全文
约1分钟内获得全文 去求助
来源期刊
CiteScore
6.10
自引率
0.00%
发文量
128
审稿时长
10 weeks
期刊最新文献
Back cover Sequence-function space of radical SAM cyclophane synthases reveal conserved active site residues that influence substrate specificity. Induced degradation of SNAP-fusion proteins. Fluorescent probes for investigating the internalisation and action of bioorthogonal ruthenium catalysts within Gram-positive bacteria. Discovery and design of molecular glue enhancers of CDK12-DDB1 interactions for targeted degradation of cyclin K.
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
现在去查看 取消
×
提示
确定
0
微信
客服QQ
Book学术公众号 扫码关注我们
反馈
×
意见反馈
请填写您的意见或建议
请填写您的手机或邮箱
已复制链接
已复制链接
快去分享给好友吧!
我知道了
×
扫码分享
扫码分享
Book学术官方微信
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术
文献互助 智能选刊 最新文献 互助须知 联系我们:info@booksci.cn
Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。
Copyright © 2023 Book学术 All rights reserved.
ghs 京公网安备 11010802042870号 京ICP备2023020795号-1