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BP: Fachverband Biologische Physik

BP 21: Poster IIIb

BP 21.34: Poster

Wednesday, March 20, 2024, 11:00–14:30, Poster C

Cell-Free Gene Expression in Bioprinted Fluidic Networks — •Alexandra Bienau and Friedrich C. Simmel — Technical University Munich, Germany

Cell-free protein expression is a valuable tool to produce specific proteins in vitro without the need for a host organism. The reduced metabolic background activity enables high product concentrations and precisely controlled reaction conditions for prototyping genetic circuitry. Microfluidic devices instead of closed reactors were used in previous works to enable longer reaction times and out-of-equilibrium behavior.

In this work, we create fluidic networks in a diffusible hydrogel environment using extrusion-based bioprinting as a fast production tool. We print sacrificial structures of Pluronics F-127 and cast agarose around them to build channels within the hydrogel, mimicking natural fluid distribution networks. The channels can be filled with customized liquids, such as the cell-free reaction mixture, and the reactions are not limited to the channels but can extend into the surrounding gel by diffusion. The behavior of fluorescent protein production within the agarose hydrogels is investigated using an E. coli-based cell extract.

Implementing gene circuitry by cell-free reactions allows for adding another design layer using bottom-up self-organization. In the future, we aim to use this design strategy to create more complex, tissue-like objects. We envision the construction of biohybrid structures combining artificial and natural cells to develop smart soft-robotic materials.

Keywords: Cell-free protein expression; Bioprinting; Fluid networks; Hydrogel; Diffusion