Cell-in-the-loop pattern formation with optogenetically emulated cell-to-cell signaling.

Area of Science:

• Synthetic biology
• Dynamical systems theory
• Biochemical engineering

Background:

• Designing synthetic biological pattern formation is complex.
• Engineering multicellular networks biochemically presents challenges.

Purpose of the Study:

• Introduce a cell-in-the-loop approach for synthetic biology.
• Develop a testbed to interrogate theoretical principles with internal cell dynamics.
• Predict and demonstrate contrasting gene expression patterns.

Main Methods:

• Utilized a cell-in-the-loop approach with in silico signaling.
• Developed a theoretical test for predicting pattern emergence.
• Emulated cell-to-cell signaling using an optogenetic setup with in silico light inputs.
• Measured real-time gene expression for feedback control.

Main Results:

• Successfully predicted spontaneous checkerboard patterns in gene expression.
• Experimentally demonstrated persistent checkerboard patterns in a 16-patch system.
• Achieved quantitative agreement between theoretical predictions and experimental results.

Conclusions:

• The cell-in-the-loop approach provides a new testbed for synthetic multicellular systems.
• Dynamical systems theory can inform the understanding of biological patterning.
• This method has potential for engineering complex synthetic multicellular systems.