L. Baldauf
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1
Expanding Through Collisions
A Dual Perspective on the Future of a Synthetic Cell
Master thesis
(2022)
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T.F. Aarts, L. Baldauf, L. van Buren, G.H. Koenderink, É. Kalmár, C. Wehrmann, M.C.A. van der Sanden
In this work, we studied future expansions of a synthetic cell with a dual perspective. For the Nanobiology part, we studied how the membrane surface area of a synthetic cell can grow, which is required for a synthetic cell to divide. Here, we combined growth strategies based on DNA and tension-mediated vesicle fusion. We found that 1 μM DNA and an osmotic shock optimise fusion efficiency, and we showcased the broad applicability of our fusion protocol. Furthermore, we observed indications of membrane growth, with a potential increase in membrane surface area sufficient for sustained synthetic cell division. For the Science Communication part, we studied the vision of the Synthetic Cell Initiative to build a European innovation ecosystem. The purpose of this innovation ecosystem is to develop synthetic cell applications through co-production. We propose that this innovation ecosystem should be formed through local synthetic cell hubs. By the promotion of bottom-up initiatives and the execution of top-down activities, responsible research and innovation can be internalised in the envisioned innovation ecosystem.
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In this work, we studied future expansions of a synthetic cell with a dual perspective. For the Nanobiology part, we studied how the membrane surface area of a synthetic cell can grow, which is required for a synthetic cell to divide. Here, we combined growth strategies based on DNA and tension-mediated vesicle fusion. We found that 1 μM DNA and an osmotic shock optimise fusion efficiency, and we showcased the broad applicability of our fusion protocol. Furthermore, we observed indications of membrane growth, with a potential increase in membrane surface area sufficient for sustained synthetic cell division. For the Science Communication part, we studied the vision of the Synthetic Cell Initiative to build a European innovation ecosystem. The purpose of this innovation ecosystem is to develop synthetic cell applications through co-production. We propose that this innovation ecosystem should be formed through local synthetic cell hubs. By the promotion of bottom-up initiatives and the execution of top-down activities, responsible research and innovation can be internalised in the envisioned innovation ecosystem.
Master thesis
(2022)
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D. de Ridder, G.H. Koenderink, G. Castro Linares, L. Baldauf, T. Idema, A. Jakobi
In animal cells, cell shape is primarily regulated by the actin cortex, a thin filament network connected to the plasma membrane. The architecture of the cortex is considered a key regulator of its function. Visualising the cortex is difficult due to the high density of numerous small-sized proteins involved in its formation. Consequently, the structure of the cortex at the membrane remains poorly studied. This study aims to gain insight into the organisation at the membrane of two key cortical components, human septin and septin-recruited actin. To study these filament structures, we reconstitute minimal cortices on supported lipid layers as model-membranes, allowing for imaging with electron and atomic force microscopy. We show that membrane binding of human septin results in ordered organisations of filaments. However, we find septin organises into arrays of paired filaments when incubated on a lipid monolayer and networks of bundles when incubated on a lipid bilayer. In addition, we showed a proof of concept for actin cortex reconstitution on lipid monolayers, which allowed us to see actin recruitment by septin meshworks.
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In animal cells, cell shape is primarily regulated by the actin cortex, a thin filament network connected to the plasma membrane. The architecture of the cortex is considered a key regulator of its function. Visualising the cortex is difficult due to the high density of numerous small-sized proteins involved in its formation. Consequently, the structure of the cortex at the membrane remains poorly studied. This study aims to gain insight into the organisation at the membrane of two key cortical components, human septin and septin-recruited actin. To study these filament structures, we reconstitute minimal cortices on supported lipid layers as model-membranes, allowing for imaging with electron and atomic force microscopy. We show that membrane binding of human septin results in ordered organisations of filaments. However, we find septin organises into arrays of paired filaments when incubated on a lipid monolayer and networks of bundles when incubated on a lipid bilayer. In addition, we showed a proof of concept for actin cortex reconstitution on lipid monolayers, which allowed us to see actin recruitment by septin meshworks.