Design and Implementation of Multi-Component Synthetic Cells

Synthetic biology has made significant strides over the past 20 years in demonstrating the ability to engineer biological systems by “programming” DNA to carry out specific operations both in cells and in cell-free systems. Currently demonstrated systems have been of modest complexity (typically less than a dozen programmed elements) and have focused on relatively simple operations (oscillators, logic operations, metabolic pathways). A major challenge in the field is learning how to systematically design and implement biomolecular circuits of much higher complexity (hundreds to thousands of programmed elements) that can carry out more complex operations, spread across multiple functional units (a la multi-cellular organisms).

Here, we will develop and demonstrate the key design tools, molecular components, and system-level architecture for synthetic cells. While we do not anticipate that a single project will lead to a functioning synthetic cell in a 3–5- year time, we believe that it will be possible implement and integrate a variety of subsystems that establish the feasibility of synthetic cells and serve as a starting point for a larger effort. The objectives for this project are to (1) develop a design-oriented, computational framework enabling modeling, analysis, and design of multi-subsystem synthetic cells, incorporating core biomolecular processes, component-level dynamics, subsystem behaviors, and multi-subsystem interactions; (2) implement and characterize a collection of biomolecular subsystems capable of providing robust and modular operations in synthetic cells; (3) demonstrate a multi-component synthetic cell capable of artificial chemotaxis using sensing of an environmental signal, decision- making based on the concentration of the environmental signal, and locomotion using a proto-flagellar motor or other similar mechanism.