Engineering of regulated stochastic cell fate determination

Area of Science:

• Synthetic biology
• Systems biology
• Cellular regulation

Background:

• Cell fate determination is crucial for development and environmental adaptation in both microbes and multicellular organisms.
• Gene regulatory networks (GRNs) control these processes, but the precise mechanisms initiating stable cell fates remain an area of active research.

Purpose of the Study:

• To engineer bistable gene networks capable of stochastic and permanent cell fate determination.
• To investigate the role of attractor basin boundaries in initiating cell fate transitions.
• To link synthetic gene networks with natural biological outputs for experimental validation.

Main Methods:

• Synthetic biology approaches to engineer bistable gene regulatory networks (GRNs).
• Integration of synthetic GRNs with natural galactose metabolism regulation in yeast.
• Mathematical modeling and computational simulations.
• Experimental validation using flow cytometry and single-cell microscopy.

Main Results:

• Demonstrated that engineered bistable GRNs can achieve permanent cell fate determination.
• Showed that inherent gene expression noise does not cause spontaneous state switching at steady state.
• Confirmed that stochastic cell fate determination requires gene expression fluctuations near attractor basin boundaries (points of instability).
• Verified model predictions through experiments with quantitatively diverse gene networks.

Conclusions:

• Rationally designed synthetic GRNs interfaced with natural regulatory mechanisms can illuminate cell differentiation principles.
• Cell differentiation and development can be initiated from points of instability within gene regulatory networks.
• This work provides insights into the intricate properties of biological networks governing cell fate decisions.