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Abstract

The bottom-up engineering of synthetic cells has emerged as a powerful field to understand the complex environment and processes of natural living cells. In particular, the development and reconstitution of synthetic cells with a cytoskeleton sets a major milestone and challenge for this aim because it is an essential, versatile and multifunctional part of all eukaryotic cells. However, current progress is limited because natural cytoskeletal components are difficult to purify, deliberately engineer and reconstitute within synthetic cells which therefore limits the otherwise multifaceted role of modern cytoskeletons. In this work, I explore means by which natural cytoskeletons can be engineered to assist the shape governing function and motility of synthetic cells. Moreover, I design synthetic cytoskeletons made from deoxyribonucleic acid (DNA) as a building block to overcome current limitations of natural cytoskeletons with DNA as a programmable and versatile tool. I show that DNA-based cytoskeletons can be reversibly assembled inside compartments with multiple stimuli, bundled into more rigid filaments, used for compartment deformation and as tracks for intracellular vesicle transport. This showcases the power of DNA cytoskeletons for bottom-up synthetic cell assembly as fully engineerable entities. All in all, I have shown how to rationally construct multifunctional natural and synthetic cytoskeletons that pave the way for engineering a synthetic cell that truly deserves its name.