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Related Concept Videos

Transcription Factors02:16

Transcription Factors

82.3K
Tissue-specific transcription factors contribute to diverse cellular functions in mammals. For example, the gene for beta globin, a major component of hemoglobin, is present in all cells of the body. However, it is only expressed in red blood cells because the transcription factors that can bind to the promoter sequences of the beta globin gene are only expressed in these cells. Tissue-specific transcription factors also ensure that mutations in these factors may impair only the function of...
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Transcription Elongation Factors02:35

Transcription Elongation Factors

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Transcription elongation is a dynamic process that alters depending upon the sequence heterogeneity of the DNA being transcribed. Hence, it is not surprising that the elongation complex's composition also varies along the way while transcribing a gene.
The transcription elongation is regulated via pausing of RNA polymerase on several occasions during transcription. In bacteria, these halts are necessary because the transcription of DNA into mRNA is coupled to the translation of that mRNA...
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Transcription Elongation Factors02:35

Transcription Elongation Factors

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Cooperative Binding of Transcription Regulators02:13

Cooperative Binding of Transcription Regulators

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Transcriptional regulators bind to specific cis-regulatory sequences in the DNA to regulate gene transcription. These cis-regulatory sequences are very short, usually less than ten nucleotide pairs in length. The short length means that there is a high probability of the exact same sequence randomly occurring throughout the genome.  Since regulators can also bind to groups of similar sequences, this further increases the chances of random binding. Transcriptional regulators form...
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Cooperative Binding of Transcription Regulators02:13

Cooperative Binding of Transcription Regulators

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General Transcription Factors01:30

General Transcription Factors

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Tissue-specific transcription factors contribute to diverse cellular functions in mammals. For example, the gene for beta globin, a major component of hemoglobin, is present in all cells of the body. However, it is only expressed in red blood cells because the transcription factors that can bind to the promoter sequences of the beta globin gene are only expressed in these cells. Tissue-specific transcription factors also ensure that mutations in these factors may impair only the function of...
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Related Experiment Video

Updated: Jan 23, 2026

High Sensitivity Measurement of Transcription Factor-DNA Binding Affinities by Competitive Titration Using Fluorescence Microscopy
06:38

High Sensitivity Measurement of Transcription Factor-DNA Binding Affinities by Competitive Titration Using Fluorescence Microscopy

Published on: February 7, 2019

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Dam mutants provide improved sensitivity and spatial resolution for profiling transcription factor binding.

Tomasz Szczesnik1,2, Joshua W K Ho1,2,3, Richard Sherwood4,5

  • 1Victor Chang Cardiac Research Institute, Darlinghurst, NSW, 2010, Australia.

Epigenetics & Chromatin
|June 15, 2019
PubMed
Summary

Modified DamID methods improve accuracy for mapping protein-DNA interactions. These advancements reduce non-specific signals and enhance spatial resolution, making it a viable alternative to ChIP-Seq for transcription factor binding studies.

Keywords:
DamDamIDTcf7l2Transcription factor

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Area of Science:

  • Molecular Biology
  • Genomics
  • Epigenetics

Background:

  • DamID is a technique for mapping protein-DNA interactions by detecting adenine methylation.
  • Current DamID methods suffer from high non-specific signals and low spatial resolution.
  • These limitations restrict DamID's application, particularly for transcription factor binding analysis.

Purpose of the Study:

  • To enhance the accuracy and resolution of the DamID technique.
  • To overcome the limitations of high non-specific signal and low spatial resolution in DamID.
  • To develop a DamID protocol suitable for sensitive transcription factor binding detection.

Main Methods:

  • Utilizing mutated Dam methylase fused to the transcription factor Tcf7l2.
  • Implementing a simplified DamID sequencing protocol.
  • Comparing the performance of modified DamID with ChIP-Seq.

Main Results:

  • Mutations in Dam significantly reduced non-specific methylation.
  • The modified DamID protocol achieved high sensitivity and spatial resolution.
  • The enhanced DamID method's performance closely matched that of ChIP-Seq.

Conclusions:

  • Modified DamID with mutated Dam methylase offers a powerful tool for mapping protein-DNA binding.
  • This improved method provides a sensitive and high-resolution alternative to ChIP-Seq.
  • The advancements enable broader applications of DamID in transcription factor research.