Sculpting and fusing biomimetic vesicle networks using optical tweezers

Abstract

Constructing higher-order vesicle assemblies has discipline-spanning potential from responsive soft-matter materials to artificial cell networks in synthetic biology. This potential is ultimately derived from the ability to compartmentalise and order chemical species in space. To unlock such applications, spatial organisation of vesicles in relation to one another must be controlled, and techniques to deliver cargo to compartments developed. Herein, we use optical tweezers to assemble, reconfigure and dismantle networks of cell-sized vesicles that, in different experimental scenarios, we engineer to exhibit several interesting properties. Vesicles are connected through double-bilayer junctions formed via electrostatically controlled adhesion. Chemically distinct vesicles are linked across length scales, from several nanometres to hundreds of micrometres, by axon-like tethers. In the former regime, patterning membranes with proteins and nanoparticles facilitates material exchange between compartments and enables laser-triggered vesicle merging. This allows us to mix and dilute content, and to initiate protein expression by delivering biomolecular reaction components.

Barcoding biological reactions with DNA-functionalized vesicles

“…[7,[67][68][69][70][71] Genetic circuitry can be harnessed in cell-free environments in order to produce av ariety of biological molecules. [7,[67][68][69][70][71] Genetic circuitry can be harnessed in cell-free environments in order to produce av ariety of biological molecules.…”

“…While previous work has shown that lipid vesicles can be engineered to fuse in order to initiate cell-free reactions, [5][6][7] the ability to rapidly and noninvasively control fusion between specific populations of vesicles,w hile limiting off-target fusion to others,h as not yet been achieved. Targeted fusion could allow for such bioreactors to be reloaded with new reagents, promoting longer durations of product synthesis.M ultiple reactions could also be simultaneously executed in the same environment with limited opportunities for reaction crosstalk.…”

Microfluidics for Biosynthesizing: from Droplets and Vesicles to Artificial Cells

“…Multicompartmentalization, a distinctive feature of living cells, can enable the separation and protection of intracellular matrix, and ensure that biological processes were protected from undesired interference. This biological characteristic has inspired scientists from multidiscipline to develop multicompartmental vesicles . In 2013, Ces and co‐workers designed a gravity‐mediated phase‐transfer process to encase multiple W/O droplets into an external bilayer, and finally fabricated the multicompartmental vesicle networks .…”

Cell‐free protein synthesis: The transition from batch reactions to minimal cells and microfluidic devices

“…Furthermore, membrane patterning, with nanoparticles and proteins, boosts the exchange of materials between compartments and also provides laser‐triggered merging of the vesicles. As a result, diluting and mixing content and commencing protein expression through the delivery of biomolecular reaction components is bestowed inside reconfigurable biomimetic networks of communicating cells (Bolognesi et al, 2018). Likewise, other forms of stimuli‐responsive artificial tissues with high levels of protein synthesis is reported.…”