Projects Bottom-up Assembly of Synthetic Cells
The evolution of cellular compartments for spatially and temporally controlled assembly of biologically processes was an essential step in developing life by evolution. Synthetic approaches to cellular-like systems are still lacking well-controlled functionalities, as would be needed for more complex synthetic cells. Towards this end, we aim to fundamentally understand how a synthetic cell with distinct cell-like functions can be constructed and, in the future, can be tailored to counter specific cellular malfunctions with pathological significance. To synthesize life’s machinery, it is indispensable to reduce the complexity of the involved systems to the fundamental. Minimization, as an important strategy in designing molecular life, can be achieved by two different approaches:
- the conventional top-down approach where components are deleted from a natural living system one after another, and
- a complementary bottom-up concept.
With the ultimate aim to construct a life-like eukaryotic cell in vitro – disentangled from the complex environment of a natural cell, we follow a bottom-up synthetic biology approach. To address this, we develop a practical and highly tunable microfluidic-based method for the high-throughput on-demand creation of synthetic cell model systems in the form of droplet-stabilized giant unilamellar vesicles (dsGUVs), polymer- and lipid-based multicompartment systems and free-standing polymersomes and GUVs.
Moreover, we design and optimize a break-through, highly controllable microfluidic technology for the sequential bottom-up assembly of more complex functional synthetic cells. This technology allows us to assemble various energy-, cytoskeleton- and adhesion-associated protein complexes within the cell-like compartments. The developed synthetic cells should be capable of self-assemble these protein complexes, and, as a consequence, generate cellular functions such as adhesion, migration and self-propelling. Beyond adhesion and motility, we aim to design autonomous life-like compartments that can produce energy and divide.
Synthetic Extracellular Vesicles for Biomedical Applications
Cell-to-cell communication is a pivotal requirement for correct functioning of multicellular organisms. The understanding of cell-to-cell communication has been revolutionized by the seminal discovery of extracellular vesicle (EV)-based intercellular signaling. EVs such as exosomes, oncosomes and microvesicles are able to shuttle genetic information between distant cells and affect nearly every facet of cell’s life, including, but not limited to migration, cell proliferation and differentiation. This broad range of EV-mediated processes also underscores their central physiological role, as well as their involvement in a wide variety of disease states. Although attaining a fundamental characterization of the intricate EV maturation and regulation mechanisms is a compelling goal for possible EVs-based therapeutic applications, the long winding and error prone procedures for EVs isolation and purification as well as their extensive complexity have hindered a full understanding.
Therefore, our primary goal is to develop a complementary and quantitative engineering approach for a sequential bottom-up assembly of fully-functional synthetic EVs with precisely-controlled lipid, protein and RNA composition. Currently, we are focusing on assembly of previously described immunology and wound healing promoting EV in their exact lipid, surface ligands and miRNA composition. Importantly, similarly to the natural wound healing promoting EVs, the assembled synthetic EVs induced pro-proliferative effects on dermal cells and increased collagen deposition as well as collective epithelial migration in human skin models. Importantly, our strategy allows to decipher the functionality of individual EV constituents. Moreover, the developed approach also opens the door for EV-based therapies that do not relay on the isolation of EVs from natural sources but rather on bio-inspired in vitro assembled fully-synthetic EVs.
These projects are part of and are funded by the MaxSynBio of the Max-Planck-Society.
