Search research articles

ABOUT JoVE

Overview Leadership Blog JoVE Help Center

AUTHORS

Publishing Process Editorial Board Scope & Policies Peer Review FAQ Submit

LIBRARIANS

Testimonials Subscriptions Access Resources Library Advisory Board FAQ

RESEARCH

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments Archive

EDUCATION

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual Faculty Resource Center Faculty Site

Terms & Conditions of Use

Related Concept Videos

Real Time RT-PCR

Real Time RT-PCR

Real-time reverse transcription-polymerase chain reaction, or Real-time RT-PCR, is an analytical tool used to determine the expression level of target genes. The method involves converting mRNA to complementary DNA with the help of an enzyme known as reverse transcriptase, followed by the PCR amplification of the cDNA. These two processes can be performed simultaneously in a single tube or separately as a two-step reaction.
The real-time quantification of the number of amplified products is...

Protein Dynamics in Living Cells

Protein Dynamics in Living Cells

Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and PET.
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by

Same author

Distinct Cdc42 protein levels differentially regulate polarized growth and cell fusion in Schizosaccharomyces pombe.

PLoS biology·2026

Same author

Basal association of a transcription factor favors early gene expression.

PLoS genetics·2025

Same author

Transcriptional heterogeneity shapes stress-adaptive responses in yeast.

Nature communications·2025

Same author

Cdc42 mobility and membrane flows regulate fission yeast cell shape and survival.

Nature communications·2024

Same author

Live cell microscopy: From image to insight.

Biophysics reviews·2024

Same author

Adapting to ever-changing conditions.

eLife·2024

Search research articles

Related Experiment Video

Updated: Jul 2, 2025

Real-time Imaging of Single Engineered RNA Transcripts in Living Cells Using Ratiometric Bimolecular Beacons

Real-time Imaging of Single Engineered RNA Transcripts in Living Cells Using Ratiometric Bimolecular Beacons

Published on: August 6, 2014

Implication of polymerase recycling for nascent transcript quantification by live cell imaging.

Olivia Kindongo¹, Guillaume Lieb¹, Benjamin Skaggs¹

¹Department of Fundamental Microbiology, Faculty of Biology and Medicine, University of Lausanne, Lausanne, Switzerland.

Yeast (Chichester, England)

|February 23, 2024

Summary

Gene length does not affect transcription intensity due to polymerase recycling. Shorter genes may produce more messenger RNA (mRNA) through this mechanism, influenced by promoter and cellular conditions.

Keywords:

MAPK signaling pathways gene looping phage‐coat proteins stress response transcription dynamics

More Related Videos

Saccharomyces cerevisiae Metabolic Labeling with 4-thiouracil and the Quantification of Newly Synthesized mRNA As a Proxy for RNA Polymerase II Activity

Saccharomyces cerevisiae Metabolic Labeling with 4-thiouracil and the Quantification of Newly Synthesized mRNA As a Proxy for RNA Polymerase II Activity

Published on: October 22, 2018

Measuring the Kinetics of mRNA Transcription in Single Living Cells

Measuring the Kinetics of mRNA Transcription in Single Living Cells

Published on: August 25, 2011

Related Experiment Videos

Last Updated: Jul 2, 2025

Real-time Imaging of Single Engineered RNA Transcripts in Living Cells Using Ratiometric Bimolecular Beacons

Real-time Imaging of Single Engineered RNA Transcripts in Living Cells Using Ratiometric Bimolecular Beacons

Published on: August 6, 2014

Saccharomyces cerevisiae Metabolic Labeling with 4-thiouracil and the Quantification of Newly Synthesized mRNA As a Proxy for RNA Polymerase II Activity

Saccharomyces cerevisiae Metabolic Labeling with 4-thiouracil and the Quantification of Newly Synthesized mRNA As a Proxy for RNA Polymerase II Activity

Published on: October 22, 2018

Measuring the Kinetics of mRNA Transcription in Single Living Cells

Measuring the Kinetics of mRNA Transcription in Single Living Cells

Published on: August 25, 2011

Area of Science:

Molecular Biology
Cell Biology
Biophysics

Background:

Live-cell imaging is essential for studying transcription dynamics.
Fluorescent probes visualize messenger RNA (mRNA) production during transcription.
Understanding factors influencing transcription signal intensity is crucial.

Purpose of the Study:

To investigate the impact of gene length on transcription site signal intensity.
To explore the mechanisms underlying transcription dynamics.
To analyze polymerase recycling in gene expression.

Main Methods:

Live-cell imaging of transcription using fluorescent probes.
Analysis of transcription signal intensity in relation to gene length.
Development and application of a mathematical model for transcription.
Experimental validation of model predictions.

Main Results:

Transcription signal intensity was independent of gene length.
A mathematical model indicated polymerase recycling explains this independence.
Experimental data confirmed that shorter genes can produce more mRNA than longer genes due to polymerase recycling.
Promoter identity and cellular state modulate polymerase recycling.

Conclusions:

Polymerase recycling significantly contributes to gene expression output.
Gene looping facilitates polymerase recycling from terminator to promoter.
The interplay between gene length, polymerase recycling, promoter, and cellular state regulates transcription.