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

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.

Automated summary

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.