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Decoupling Resource-Coupled Gene Expression in Living Cells.

Tatenda Shopera1, Lian He1, Tolutola Oyetunde1

  • 1Department of Energy, Environmental and Chemical Engineering, Washington University in St. Louis , St. Louis, Missouri 63130, United States.

ACS Synthetic Biology
|May 2, 2017
PubMed
Summary
This summary is machine-generated.

Synthetic biology aims to build complex gene networks. This study shows negative feedback reduces resource interference between genetic circuits, enabling reliable multitasking in cells.

Keywords:
biological robustnessnegative feedbackprotein sequestrationregulatory architectureresource competition

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Area of Science:

  • Synthetic biology
  • Systems biology
  • Metabolic engineering

Background:

  • Synthetic biology seeks to engineer reliable gene networks with predictable functions.
  • Increasing network complexity strains cellular resources, causing interference between non-interacting circuits.
  • Mechanisms to mitigate resource competition in complex genetic systems are not well understood.

Purpose of the Study:

  • To investigate resource-associated interference in synthetic gene networks.
  • To determine if negative feedback can decouple interacting genetic circuits.
  • To reveal design principles for reliable multitasking in engineered cells.

Main Methods:

  • Construction of three Escherichia coli systems with two independent circuit modules.
  • Systematic increase in the complexity of Circuit 2 while Circuit 1 remained constant.
  • Computational modeling and experimental validation of circuit behaviors and interference.

Main Results:

  • Demonstrated indirect coupling between seemingly independent genetic circuits across three regulatory topologies.
  • Verified computationally and experimentally that negative feedback significantly reduces resource-coupled interference.
  • Observed that engineered negative feedback enhances the ability of cells to control cellular resources.

Conclusions:

  • Indirect coupling due to resource competition is a significant challenge in complex synthetic gene networks.
  • Negative feedback serves as a key design principle to decouple genetic circuits and minimize interference.
  • This work provides a strategy for engineering robust cellular systems capable of reliable multitasking.