Real-Time Imaging of Single Viral mRNA Translation in Live Cells Using CRISPR/dCas13.
Real-Time Imaging of Single Viral mRNA Translation in Live Cells Using CRISPR/dCas13.
复制标题
DOI：
10.1021/acs.analchem.3c03365
复制
发表时间：
2023-10
影响因子：
7.4
通讯作者：
Yi-Fan Tian;Yu-Peng Zhang;Qiu-Mei Wu;Dai-Wen Pang;Shu-Lin Liu;Zhi-Gang Wang
中科院分区：
文献类型：
--
作者：
Yi-Fan Tian;Yu-Peng Zhang;Qiu-Mei Wu;Dai-Wen Pang;Shu-Lin Liu;Zhi-Gang Wang
翻译是病毒入侵宿主细胞后调节的许多关键细胞活动之一，病毒mRNA翻译动力学和亚细胞定位的研究需要动态，实时可视化的翻译技术。然而，用于成像mRNA翻译的常规工具通常需要可能影响天然翻译的编码区修饰。在这里，我们使用一种工具实现了翻译的动态成像，该工具使用具有特定互补配对指导RNA的CRISPR/dCas 13系统标记具有未修饰编码区的靶mRNA。该系统实现了翻译过程的实时动态可视化，是进一步研究翻译机制的一个有前途的工具。
Translation is one of the many critical cellular activities regulated by viruses following host-cell invasion, and studies of viral mRNA translation kinetics and subcellular localization require techniques for the dynamic, real-time visualization of translation. However, conventional tools for imaging mRNA translation often require coding region modifications that may affect native translation. Here, we achieve dynamic imaging of translation with a tool that labels target mRNAs with unmodified coding regions using a CRISPR/dCas13 system with specific complementary paired guide RNAs. This system enables a real-time dynamic visualization of the translation process and is a promising tool for further investigations of the mechanisms of translation.