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Related Concept Videos

Combinatorial Gene Control02:33

Combinatorial Gene Control

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Combinatorial gene control is the synergistic action of several transcriptional factors to regulate the expression of a single gene. The absence of one or more of these factors may lead to a significant difference in the level of gene expression or repression.
The expression of more than 30,000 genes is controlled by approximately 2000-3000 transcription factors. This is possible because a single transcription factor can recognize more than one regulatory sequence. The specificity in gene...
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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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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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Updated: Jun 11, 2025

Rapid Synthesis and Screening of Chemically Activated Transcription Factors with GFP-based Reporters
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An Automated Cell-Free Workflow for Transcription Factor Engineering.

Holly M Ekas1,2,3, Brenda Wang1,2,3, Adam D Silverman1,2,3

  • 1Department of Chemical and Biological Engineering, Northwestern University, Evanston, Illinois 60208, United States.

ACS Synthetic Biology
|October 7, 2024
PubMed
Summary
This summary is machine-generated.

We developed a rapid, automated cell-free gene expression (CFE) system using acoustic liquid handling. This accelerates the engineering of biosensors for detecting heavy metals like mercury and cadmium.

Keywords:
cell-free gene expressionhigh-throughputprotein engineeringrobotic liquid handlingsynthetic biologytranscription factor

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Area of Science:

  • Synthetic Biology
  • Biotechnology
  • Biochemistry

Background:

  • High-throughput assays are crucial for optimizing biological systems but are often slow to develop.
  • Cell-free gene expression (CFE) offers advantages over in vivo assays for flexibility and speed.
  • Automated platforms are needed to streamline the assay development process.

Purpose of the Study:

  • To create a generalizable and optimized automated cell-free gene expression (CFE) workflow.
  • To accelerate the design-build-test-learn cycles in synthetic biology.
  • To engineer transcription factor-based biosensors for enhanced metal detection.

Main Methods:

  • Developed a generalizable approach for optimizing automated CFE workflows using the Echo Acoustic Liquid Handler.
  • Implemented pilot assays and validation strategies for protocol development.
  • Engineered transcription factor variants (MerR and CadR) for mercury and cadmium detection.

Main Results:

  • Rapidly generated and assayed 127 MerR and 134 CadR variants in 3682 CFE reactions within 48 hours.
  • Improved biosensor limit of detection, selectivity, and dynamic range for mercury and cadmium.
  • Assessed sensitivity and selectivity against a panel of metal ions.

Conclusions:

  • The Echo-based, automated CFE platform significantly accelerates the development of engineered biosensors.
  • This approach can be broadly applied to various design workflows in synthetic biology.
  • Enables rapid optimization of transcription factor variants for specific analyte detection.