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Related Concept Videos

Protein Dynamics in Living Cells01:19

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...
Real Time RT-PCR02:57

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...

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Related Experiment Video

Updated: May 8, 2026

Measuring the Kinetics of mRNA Transcription in Single Living Cells
11:22

Measuring the Kinetics of mRNA Transcription in Single Living Cells

Published on: August 25, 2011

Measuring transcription dynamics in living cells using fluctuation analysis.

Matthew L Ferguson1, Daniel R Larson

  • 1Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA.

Methods in Molecular Biology (Clifton, N.J.)
|August 28, 2013
PubMed
Summary

Researchers developed a new method to visualize and quantify single gene transcription in living cells. This technique uses fluorescent proteins and RNA hairpins to track pre-mRNA synthesis, enabling detailed analysis of transcription dynamics.

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Real-time Analysis of Transcription Factor Binding, Transcription, Translation, and Turnover to Display Global Events During Cellular Activation
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Last Updated: May 8, 2026

Measuring the Kinetics of mRNA Transcription in Single Living Cells
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Published on: August 25, 2011

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Real-time Analysis of Transcription Factor Binding, Transcription, Translation, and Turnover to Display Global Events During Cellular Activation
12:54

Real-time Analysis of Transcription Factor Binding, Transcription, Translation, and Turnover to Display Global Events During Cellular Activation

Published on: March 7, 2018

Area of Science:

  • Molecular Biology
  • Cell Biology
  • Genetics

Background:

  • Single-cell studies reveal transcription dynamics and nuclear organization influence gene expression heterogeneity.
  • Understanding transcriptional regulation requires high-resolution methods for observing gene activity in real-time.

Purpose of the Study:

  • To describe a novel approach for visualizing and quantifying pre-messenger RNA (pre-mRNA) synthesis at single active genes in living cells.
  • To enable real-time observation of transcription with high spatial and temporal resolution.

Main Methods:

  • Utilizes the high-affinity interaction between MS2/PP7 bacteriophage coat proteins and specific RNA hairpins transcribed by the gene of interest.
  • Employs MS2/PP7 coat proteins fused to fluorescent proteins to label nascent mRNA for detection via fluorescence microscopy.
  • Employs time-lapse fluorescence imaging and quantitative image analysis to generate fluorescence intensity time traces at transcription sites.
  • Applies temporal autocorrelation analysis to determine RNA polymerase (RNAP) enzymatic activities, including initiation and elongation rates.

Main Results:

  • Successfully visualized and quantified pre-mRNA synthesis at single active genes in real-time within living cells.
  • Generated detailed time traces of fluorescence intensity at transcription sites.
  • Quantified key RNAP enzymatic activities, such as initiation and elongation rates, through temporal autocorrelation analysis.

Conclusions:

  • The described approach provides a powerful tool for real-time observation and quantitative analysis of transcription dynamics at the single-gene level in living cells.
  • This method facilitates a holistic understanding of transcriptional regulation by integrating gene activity with spatial and temporal cellular contexts.