A Covalent Approach for Site-Specific RNA Labeling in Mammalian Cells

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• 英文论文题名：A Covalent Approach for Site-Specific RNA Labeling in Mammalian Cells
• 论文作者：Fahui Li ; Jianshu Dong ; Xiaosong Hu ; Weimin Gong ; Jiasong Li ; Jing Shen ; Huifang Tian ; 王江云
• 英文论文作者：Fahui Li ; Jianshu Dong ; Xiaosong Hu ; Weimin Gong ; Jiasong Li ; Jing Shen ; Huifang Tian ; Jiangyun Wang ; Institute of Biophysics ; Chinese Academy of Sciences
• 年：2016
• 作者机构：中国科学院生物物理研究所;
• 英文论文关键词：bioorthogonal chemistry ; biotechnology ; RNA labeling ; RNA modification ; tRNA
• 会议召开时间：2016-09-25
• 会议录名称：第九届全国核糖核酸学术讨论会论文集
• 语种：英文
• 分类号：Q953
• 学会代码：IGSS
• 会议名称：第九届全国核糖核酸学术讨论会
• 会议地点：中国上海
• 主办单位：中国生物化学与分子生物学会核糖核酸专业委员会
• 学会名称：中国生物化学与分子生物学会
• 页数：1
• 文件大小：385k
• 原文格式：D
• 会议级别：全国

摘要

Advances in RNA research and RNA nanotechnology depend on the ability to manipulate and probe RNA with high precision through chemical approaches,both in vitro and in mammalian cells.However,covalentRNA labeling methods with scope and versatility comparable to those of current protein labeling strategies are underdeveloped.A method is reported for the site-and sequence-specific covalent labeling of RNAs in mammalian cells by using tRNAIle2-agmatidine synthetase(Tias) and click chemistry.The crystal structure of Tias in complex with an azide-bearing agmatine analogue was solved to unravel the structural basis for Tias/substrate recognition.The unique RNA sequence specificity and plastic Tias/substrate recognition enable the site-specific transfer of azide/alkyne groups to an RNA molecule of interest in vitro and in mammalian cells.Subsequent click chemistry reactions facilitate the versatile labeling,functionalization,and visualization of targetRNA.
Advances in RNA research and RNA nanotechnology depend on the ability to manipulate and probe RNA with high precision through chemical approaches,both in vitro and in mammalian cells.However,covalentRNA labeling methods with scope and versatility comparable to those of current protein labeling strategies are underdeveloped.A method is reported for the site- and sequence-specific covalent labeling of RNAs in mammalian cells by using tRNAIle2-agmatidine synthetase(Tias) and click chemistry.The crystal structure of Tias in complex with an azide-bearing agmatine analogue was solved to unravel the structural basis for Tias/substrate recognition.The unique RNA sequence specificity and plastic Tias/substrate recognition enable the site-specific transfer of azide/alkyne groups to an RNA molecule of interest in vitro and in mammalian cells.Subsequent click chemistry reactions facilitate the versatile labeling,functionalization,and visualization of targetRNA.

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