Real-Time Imaging of Single Viral mRNA Translation in Live Cells Using CRISPR/dCas13,Analytical Chemistry

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Real-Time Imaging of Single Viral mRNA Translation in Live Cells Using CRISPR/dCas13
Analytical Chemistry ( IF 6.7 ) Pub Date : 2023-10-24 , DOI: 10.1021/acs.analchem.3c03365
Yi-Fan Tian ₁ , Yu-Peng Zhang ₁ , Qiu-Mei Wu ₂ , Dai-Wen Pang ₁ , Shu-Lin Liu ₁ , Zhi-Gang Wang ₁

Affiliation

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.

中文翻译：

使用 CRISPR/dCas13 对活细胞中单个病毒 mRNA 翻译进行实时成像

翻译是宿主细胞入侵后受病毒调节的众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、实时翻译可视化技术。然而，用于成像 mRNA 翻译的传统工具通常需要编码区修改，这可能会影响天然翻译。在这里，我们使用带有特定互补配对向导RNA的CRISPR/dCas13系统来标记具有未修饰编码区的目标mRNA的工具来实现翻译的动态成像。该系统可以实现翻译过程的实时动态可视化，是进一步研究翻译机制的有前途的工具。

更新日期：2023-10-24

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