A novel reaction-diffusion architecture for engineering self-organized patterns in mammalian cells

Area of Science:

• Synthetic biology
• Developmental biology
• Biophysics

Background:

• Reaction-diffusion systems create spatial patterns via local activation and long-range inhibition.
• Synthetic implementations in mammalian cells are hindered by the need for differential diffusion rates.

Purpose of the Study:

• Introduce and validate a novel synthetic biology architecture, juxtacrine activation with paracrine inhibition (JAPI).
• Demonstrate JAPI's capability for self-organized multicellular patterning in mammalian cells.
• Explore JAPI's potential for programming complex spatial patterns and developmental processes.

Main Methods:

• Mathematical and numerical modeling of the JAPI system.
• Engineering compact synNotch-based JAPI circuits in mammalian fibroblasts.
• Perturbing feather bud formation in embryonic chicken epidermis using JAPI circuits.
• Developing a library-based approach for coupled, dual-JAPI circuits.

Main Results:

• JAPI system mathematically and numerically shown to access patterning regimes similar to diffusion-based circuits with fewer parameters.
• Engineered JAPI circuits demonstrate tunable, size-limited signal propagation for self-organized patterning.
• JAPI circuits successfully perturbed feather bud development in vivo.
• Coupled dual-JAPI circuits enable programmable pattern interactions and access to diverse spatial states.

Conclusions:

• JAPI offers a novel, compact, and modular platform for programming self-organized multicellular patterning.
• This approach overcomes limitations of previous synthetic reaction-diffusion systems.
• JAPI has significant potential for applications in developmental biology and synthetic biology.