The chassis
Our group has pioneered experiments to reconstitute the flow of genetic information (DNA → mRNA → protein), a universal process in every organism, inside liposomes. We use PURE (Protein synthesis Using Recombinant Elements) system as a minimal gene expression platform. We are able to produce tens of thousands of gene-expressing liposomes per <10-μL samples and perform high-content fluorescence microscopy and (imaging) flow cytometry.
Building the modules
DNA replication
We created the first autocatalytic network in PURE system whereby a self-replicating DNA expresses proteins that catalyse its own amplification, forming a positive feedback loop. We drew inspiration from the protein-primed DNA replication mechanism of the phi29 virus.
Membrane remodelling & division
The canonical pathway for bacterial division involves the membrane constricting Z-ring. We reconstituted the basal Z-ring proteins (FtsA and FtsZ) in PURE system. Membrane necks and budding vesicles are formed, sometimes generating dumbbell-shape liposomes. This is the first example of genetically encoded liposome constriction.
Min oscillations
The Min system has been functionally expressed inside liposomes, creating oscillatory patterns induced by the dynamic self-organisation of Min proteins in the vesicle lumen and the membrane. Periodic liposome deformation events driven by Min redistribution have been observed, showcasing the unexpected behaviour of some reconstituted protein systems.
Lipid synthesis
Homeostatic growth of the cellular membrane is a hallmark of free-living organisms. Phospholipids, the main constituents of biological membranes have been synthesised in PURE system by expressing all seven enzymes of the E. coli Kennedy pathway.
Module integration & Evolution
We are engineering synthetic cells via multiple rounds of module integration and evolutionary optimisation. We use an integrative approach combining de novo design and assembly of synthetic genomes, genetic diversification, high-throughput screening methods, next-generation DNA sequencing, automated microscopy, AI-assisted image/data analysis, and system's level evolution. Our primary goal is the integration of DNA replication, evolvability (self-improvement), regeneration of PURE translation factors, membrane synthesis, and liposome division from a synthetic genome. Read our roadmap article towards autonomy in synthetic cells.
Synthetic cell technologies
We develop foundational technologies for the production of tailored synthetic cells. Specifically, we develop AI-powered lab automation to enhance PURE expression and a suite of technological innovations to scale up the production of liposomes and select those with the desired properties. We aim at creating the first synthetic cell foundry as a solution to apply synthetic cells in biotechnology and personalised medicine.