Related Experiment Video
Updated: Jul 2, 2025
12:20
Real-time Imaging of Single Engineered RNA Transcripts in Living Cells Using Ratiometric Bimolecular Beacons
Published on: August 6, 2014
11.8K
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.
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.
Keywords:
MAPK signaling pathwaysgene loopingphage‐coat proteinsstress responsetranscription dynamicsMore Related Videos
Related Concept Videos
02:57Real Time RT-PCR
57.3K
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...
The real-time quantification of the number of amplified products is...
57.3K
01:19Protein Dynamics in Living Cells
2.1K
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...
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...
2.1K