Quantifying tagged mRNA export flux via nuclear pore complexes in single live cells

Yueyue Jing; Yilin Lv; Jingya Ye; Longfang Yao; Liwen Chen; Lan Mi; Yiyan Fei; Yao Yu; Biao Dong; Hong Lv; Jiong Ma

doi:10.1016/j.bbrc.2021.01.049

Quantifying tagged mRNA export flux via nuclear pore complexes in single live cells

Biochem Biophys Res Commun. 2021 Mar 19:545:138-144. doi: 10.1016/j.bbrc.2021.01.049. Epub 2021 Feb 3.

Authors

Yueyue Jing¹, Yilin Lv², Jingya Ye³, Longfang Yao¹, Liwen Chen¹, Lan Mi¹, Yiyan Fei¹, Yao Yu², Biao Dong⁴, Hong Lv⁵, Jiong Ma⁶

Affiliations

¹ Department of Optical Science and Engineering, Shanghai Engineering Research Center of Ultra-Precision Optical Manufacturing, Key Laboratory of Micro and Nano Photonic Structures (Ministry of Education), Green Photoelectron Platform, Fudan University, Shanghai, China.
² State Key Laboratory of Genetic Engineering, School of Life Sciences, Fudan University, Shanghai, 200438, China; Shanghai Engineering Research Center of Industrial Microorganisms, Shanghai, 200438, China.
³ National Clinical Research Center for Geriatrics, West China Hospital, Sichuan University, Sichuan, 610041, China.
⁴ National Clinical Research Center for Geriatrics, West China Hospital, Sichuan University, Sichuan, 610041, China. Electronic address: biaodong@scu.edu.cn.
⁵ State Key Laboratory of Genetic Engineering, School of Life Sciences, Fudan University, Shanghai, 200438, China; Shanghai Engineering Research Center of Industrial Microorganisms, Shanghai, 200438, China. Electronic address: honglv@fudan.edu.cn.
⁶ Department of Optical Science and Engineering, Shanghai Engineering Research Center of Ultra-Precision Optical Manufacturing, Key Laboratory of Micro and Nano Photonic Structures (Ministry of Education), Green Photoelectron Platform, Fudan University, Shanghai, China; Institute of Biomedical Engineering and Technology, Academy for Engineer and Technology, Fudan University, Shanghai, China; Shanghai Engineering Research Center of Industrial Microorganisms, Multiscale Research Institute of Complex Systems (MRICS), School of Life Sciences, Fudan University, Shanghai, China. Electronic address: jiongma@fudan.edu.cn.

PMID: 33548627
DOI: 10.1016/j.bbrc.2021.01.049

Abstract

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.

Keywords: Fluorescence microscopy; Gene expression monitoring; Modeling; Nuclear pore; RNA Transport.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Active Transport, Cell Nucleus*
Catabolite Repression
Dependovirus / genetics
Dependovirus / metabolism
Gene Expression
Genes, Fungal
HeLa Cells
Humans
Kluyveromyces / genetics
Kluyveromyces / metabolism
Luminescent Proteins / genetics
Luminescent Proteins / metabolism
Models, Biological
Nuclear Pore / metabolism*
RNA, Fungal / genetics
RNA, Fungal / metabolism
RNA, Messenger / genetics
RNA, Messenger / metabolism*
RNA, Viral / genetics
RNA, Viral / metabolism
Red Fluorescent Protein
Single-Cell Analysis
Transfection

Substances

Luminescent Proteins
RNA, Fungal
RNA, Messenger
RNA, Viral

Supplementary concepts

Kluyveromyces marxianus