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Related Concept Videos

Combinatorial Gene Control02:33

Combinatorial Gene Control

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...
Translesion DNA Polymerases02:10

Translesion DNA Polymerases

Translesion (TLS) polymerases rescue stalled DNA polymerases at sites of damaged bases by replacing the replicative polymerase and installing a nucleotide across the damaged site. Doing so, TLS allows additional time for the cell to repair the damage before resuming regular DNA replication.
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Regulation of Expression at Multiple Steps

The gene expression in cells is regulated at different stages: (i) transcription, (ii) RNA processing, (iii) RNA localization, and (iv) translation. Transcriptional regulation is mediated by regulatory proteins such as transcription factors, activators, or repressors—these control gene expression by initiating or inhibiting the transcription of genes. Once a precursor or pre-mRNA is produced, it undergoes post-transcriptional modification, including 5' capping, splicing, and the addition of a...
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The operon model represents a fundamental mechanism of gene regulation in prokaryotes, enabling coordinated expression of genes involved in related metabolic or functional pathways. Operons consist of structural genes, a promoter, and an operator, with transcription regulated by repressors, activators, and small effector molecules.Structure and Function of OperonsAn operon is a cluster of structural genes transcribed together under the control of a single promoter. The promoter region...
Conservative Site-specific Recombination and Phase Variation02:53

Conservative Site-specific Recombination and Phase Variation

Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
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Rapid Assembly of Multi-Gene Constructs using Modular Golden Gate Cloning
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Modular control of multiple pathways using engineered orthogonal T7 polymerases.

Karsten Temme1, Rena Hill, Thomas H Segall-Shapiro

  • 1UCB/UCSF Joint Graduate Group in Bioengineering, MC2540, Department of Pharmaceutical Chemistry, School of Pharmacy, University of California, San Francisco, 1700 4th Street, San Francisco, CA 94158, USA.

Nucleic Acids Research
|June 30, 2012
PubMed
Summary
This summary is machine-generated.

This study introduces a novel synthetic biology design separating genetic circuits from gene pathways using engineered polymerases. This system allows precise, programmable control over gene expression and cellular functions.

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Area of Science:

  • Synthetic biology
  • Genetic engineering
  • Systems biology

Background:

  • Synthetic genetic circuits offer programmable control over gene expression for complex cellular functions.
  • Integrating circuits with target pathways can be challenging, potentially leading to toxicity or evolutionary pressure.

Purpose of the Study:

  • To develop a design strategy for genetically separating sensing/circuitry functions from controlled gene pathways.
  • To create a modular system for precise, orthogonal control of multiple gene pathways.

Main Methods:

  • Engineered T7 RNA polymerase variants to create four orthogonal polymerases.
  • Designed a 'controller' plasmid encoding sensors, circuits, and polymerases.
  • Utilized polymerase-specific promoters for pathway activation.
  • Demonstrated functionality with a controller integrating two inducible systems and AND logic.

Main Results:

  • Developed four highly orthogonal polymerases with minimal off-target promoter activity (8- to 75-fold induction).
  • Successfully demonstrated independent control channels for linking circuit outputs to cellular functions.
  • Implemented a controller plasmid enabling logic operations and switching between metabolic pathways (producing green and red pigments).

Conclusions:

  • The proposed design strategy enables modular and tunable control of gene expression by separating regulatory circuits from gene pathways.
  • This approach enhances orthogonality, reduces pathway gene expression in the absence of a controller, and simplifies pathway regulation changes.
  • The system offers advantages for managing complex synthetic biology applications and metabolic engineering in organisms like Escherichia coli.