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

被引:2
作者
Tian, Yi-Fan [1 ,2 ]
Zhang, Yu-Peng [1 ,2 ]
Wu, Qiu-Mei [3 ]
Pang, Dai-Wen [1 ,2 ]
Liu, Shu-Lin [1 ,2 ]
Wang, Zhi-Gang [1 ,2 ]
机构
[1] Nankai Univ, State Key Lab Med Chem Biol, Tianjin Key Lab Biosensing & Mol Recognit, Res Ctr Analyt Sci,Coll Chem, Tianjin, Peoples R China
[2] Nankai Univ, Sch Med, Tianjin, Peoples R China
[3] Fujian Med Univ, Sch Basic Med Sci, Minist Educ, Key Lab Gastrointestinal Canc, Fuzhou 350122, Peoples R China
基金
中国国家自然科学基金;
关键词
LIVING CELLS; DYNAMICS; TRACKING; PROTEINS;
D O I
10.1021/acs.analchem.3c03365
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
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.
引用
收藏
页码:16298 / 16304
页数:7
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