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Area of Science:

  • Synthetic Biology
  • Cellular Engineering
  • Microbial Ecology

Background:

  • Cellular engineering faces challenges in heterogeneous environments due to unpredictable cellular responses.
  • Non-uniform nutrient availability drives complex cell-environment interactions and phenotypic differentiation.
  • Controlling spatial gene expression in synthetic biology remains difficult under non-uniform conditions.

Purpose of the Study:

  • To design gene circuits that sense and control phenotypic structure in bacterial microcolonies.
  • To leverage Escherichia coli's stress response for growth arrest sensing and phenotypic control.
  • To develop strategies for synthetic biology applications in complex, heterogeneous environments.

Main Methods:

  • Designed gene circuits coupling tunable sensors to actuator and inducible gating modules.
  • Utilized Escherichia coli's stress response activated on growth arrest.
  • Implemented environmental feedback loops for robust growth-dormancy cycling.
  • Employed a spatiotemporal computational model for prediction and validation.
  • Developed a 'stress-gated lysis circuit' to eliminate dormant phenotypes.

Main Results:

  • Demonstrated robust cycling between growth and dormancy in bacterial microcolonies via environmental feedback.
  • Successfully controlled population structure by selectively eliminating dormant cells.
  • Validated circuit behavior using a spatiotemporal computational model.
  • Established a method to modulate microbial colony structure.

Conclusions:

  • Developed a novel strategy to sense and control bacterial phenotypic structure in microcolonies.
  • Showcased the potential of engineered gene circuits for applications in complex environments.
  • Provided a framework for leveraging environmental feedback to manage microbial populations.