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Related Concept Videos

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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Transcription Factors02:16

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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Transcription01:10

Transcription

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Overview
Transcription is the process of synthesizing RNA from a DNA sequence by RNA polymerase. It is the first step in producing a protein from a gene sequence. Additionally, many other proteins and regulatory sequences are involved in the proper synthesis of messenger RNA (mRNA). Regulation of transcription is responsible for the differentiation of all the different types of cells and often for the proper cellular response to environmental signals.
Transcription Can Produce Different Kinds...
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Master Transcription Regulators02:23

Master Transcription Regulators

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Master transcription regulators are regulatory proteins that are predominantly responsible for regulating the expression of multiple genes. Often these genes work in concert to drive a  complex process. Activation of a master transcription regulator can lead to a cascade of transcriptional activation necessary for that outcome. These regulators can directly bind to the regulatory sequences of the various genes involved, or they can indirectly regulate transcription by binding to regulatory...
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Eukaryotic Transcription Inhibitors01:52

Eukaryotic Transcription Inhibitors

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Certain biochemical processes, such as embryonic development and cell growth regulation, depend on the repression of specific genes. DNA binding proteins known as eukaryotic transcription inhibitors regulate the repression of gene expression in eukaryotes. The presence of these inhibitors at the required location and time in the cell is triggered by the presence of hormones and additional signals from other cells.
Eukaryotic transcription inhibitors usually contain two distinct domains, a...
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Related Experiment Video

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Evaluation of a Universal Nested Reverse Transcription Polymerase Chain Reaction for the Detection of Lyssaviruses
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Evaluation of a Universal Nested Reverse Transcription Polymerase Chain Reaction for the Detection of Lyssaviruses

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Versatile transcription control based on reversible dCas9 binding.

Julia R Widom1,2, Victoria Rai1,2,3, Christopher E Rohlman4

  • 1Single Molecule Analysis Group, Department of Chemistry, University of Michigan, Ann Arbor, Michigan 48109, USA.

RNA (New York, N.Y.)
|July 20, 2019
PubMed
Summary
This summary is machine-generated.

This study introduces a novel CRISPR-Cas9 system for precise, time-controlled RNA production in vitro. This method allows independent control over multiple RNA transcripts, advancing molecular biology and nanotechnology applications.

Keywords:
CRISPRRNA polymerasereversibilitytranscription

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

  • Molecular Biology
  • Synthetic Biology
  • Biotechnology

Background:

  • Controlling transcription in vitro is crucial for molecular biology and nanotechnology.
  • Existing methods lack precise, independent control over multiple RNA transcript production.
  • CRISPR-Cas9 technology offers potential for targeted DNA manipulation.

Purpose of the Study:

  • To develop a method for precise, time-dependent control of multiple RNA transcript production in vitro.
  • To utilize catalytically dead Streptococcus pyogenes CRISPR-Cas9 (dCas9) and single guide RNA (sgRNA) for transcription blockade.
  • To enable on-demand activation of specific RNA synthesis.

Main Methods:

  • Employing catalytically dead Streptococcus pyogenes CRISPR-Cas9 (dCas9) complexed with sgRNA to create transcription blockades on DNA templates.
  • Testing blockade efficiency against RNA polymerases (RNAPs) from bacteriophages SP6, T3, T7, and Escherichia coli.
  • Investigating blockade robustness with mismatched sgRNA sequences and on-demand activation using competitor DNA.

Main Results:

  • dCas9:sgRNA complexes achieved >99.5% blockade efficiency against SP6, T3, and T7 RNAPs, causing dissociation.
  • A ~70% blockade efficiency was observed for Escherichia coli RNAP, with sustained binding.
  • Blockade efficiency remained >95% with four mismatches in the sgRNA target sequence, enabling competitor-DNA-mediated activation.

Conclusions:

  • The dCas9:sgRNA system provides robust and efficient transcription blockade in vitro.
  • This strategy allows for precise, independent, and temporally controlled production of multiple RNA species from a single DNA template.
  • The developed method represents a significant advancement for transcription control in synthetic biology and nanotechnology.