Bottom-up synthetic biology has been successful at isolating components from cells and reconstituting subcellular functions inside compartments. Progress towards a fully functional synthetic cell, however, requires strategies to recombine and arrange a multitude of components in space and time. We therefore propose to merge two precision technologies, namely microfluidics and DNA nanotechnology, to position and manipulate components in synthetic cells[1,2]. In particular, we demonstrate that DNA can be used as a near-universal linkerfor responsive and programmable compartment functionali-zation. Our method relies on the self-assembly of single-stranded cholesterol-tagged DNA handles, which provide an addressable anchoring point for complementary DNA carrying an arbitrary functional group. Using this DNA handle approach, we demonstrate the stimuli-responsive attachment of reactive groups, DNA nanostructures, microspheres, an actin cortex and even living cells to the periphery of surfactant-stabilized microfluidic droplets[3]. We further demonstrate a simple one-pot method for the assembly of complex giant unilamellar vesicle (GUV)-based syntheticcells[4]. Within the GUVs, we employ DNA to construct functional components, including a pH-responsive DNA-based cytoskeleton mimic, which serves as a stabilizing cortex.
[1]K. Göpfrich*, I. Platzman* & J. P. Spatz*, Mastering Complexity: Towards Bottom-up Construction of Multifunctional Eukaryotic Synthetic Cells, Trends in Biotechnology36, 938–951(2018).
[2] B. Haller, K. Göpfrich, M. Schröter, J. W. Janiesch, I. Platzman* & J. P. Spatz*, Charge-controlled microfluidic formation of lipid-based single-and multicompartment systems, Lab on a Chip18, 2665–2674(2018).
[3]K. Jahnke, M. Weiss, C. Frey, S. Antona, J.-W. Janiesch, I. Platzman, K. Göpfrich* & J. P. Spatz*, Programmable Functionalization of Surfactant-Stabilized Microfluidic Droplets via DNA-Tags, Advanced Functional Materials(2019).doi: 10.1002/adfm.201808647.
[4]K. Göpfrich, B. Haller,O. Staufer, Y. Dreher, U. Mehrsdorf, I. Platzman*& J. P. Spatz*, One-Pot Assembly of Complex Giant Unilamellar Vesicle-BasedSynthetic Cells, ACS Synthetic Biology, 8, 937–947(2019). doi: 10.1021/acssynbio.9b00034.