Journal
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
Volume 545, Issue -, Pages 138-144Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.bbrc.2021.01.049
Keywords
Gene expression monitoring; RNA Transport; Nuclear pore; Fluorescence microscopy; Modeling
Categories
Funding
- National Natural Science Foundation of China (NSFC) [61575046, 11574056, 81770199]
- National Key R&D Program of China [2016YFC0905200]
- Ministry of Science and Technology of the People's Republic of China, the China-Serbia bilateral project [SINO-SERBIA2018002]
- Fudan University-CIOMP Joint Fund [FC2017-007, FC2018-001]
- Pioneering Project of Academy for Engineering and Technology, Fudan University [gyy2018-001, gyy2018-002]
- Shanghai Science and Technology Commission [19DZ2282100]
- Shanghai Natural Science Foundation [20ZR1405100, 20ZR1403700]
- Scientific Promotion Program of Outstanding Postgraduate, Fudan University [SSH6281011]
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A new method has been developed to measure the export flux of mRNA in single live cells using snapshot images, allowing for direct monitoring of gene behavior and understanding of gene regulation. The technique has been validated in HeLa cells and yeast cells, showing its noninvasive nature and potential for research in gene expression.
The mRNA export flux through nuclear pore complexes (NPC) changes under DNA manipulation and hence affects protein translation. However, monitoring the flux of a specific mRNA in single live cell is beyond reach of traditional techniques. We developed a fluorescence-based detection method for measuring the export flux of mRNA through NPC in single live cell using a snapshot image, which had been tested on exogenous genes' expression in HeLa cells, with transfection or infection, and endogenous genes' expression in yeast cells, during incubation and carbon catabolite repression. With its speediness, explicitness and noninvasiveness, we believe that it would be valuable in direct monitoring of gene behavior, and the understanding of gene regulation at a single cell level. (c) 2021 Elsevier Inc. All rights reserved.
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