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From pattern formation of cell-division proteins in shaped bacteria towards bottom up assembly of a synthetic divisome

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If you have a question about this talk, please contact Dr. Ulrich Keyser.

In my group we use the tools of nanotechnology (nanofabrication, tweezers, fast AFM , fluorescence…) to explore biology at the single-molecule and single-cell scale. Our research ranges from single-molecule biophysics studies of DNA -protein interactions to DNA translocation through solid-state nanopores to exploring biophysics of bacteria with nanofabricated shapes, see http://ceesdekkerlab.tudelft.nl

Recently we have begun a project aimed at realizing synthetic cell division [1]. We work towards building liposomes (lipid vesicles enclosing an aqueous solution with purified proteins and DNA ) that can spontaneously divide through a contractile protein ring at the vesicle’s perimeter. To realize this, we employ an experimental biophysics approach that addresses both the actual division and the prerequisite spatial control, studying: (i) Cells in nanofabricated shapes. We study cell-division proteins and DNA in live E.coli bacteria that are molded into user-defined arbitrary shapes and sizes. Clarifying the effects of cell shape will elucidate the guiding principles for the spatiotemporal organization of the cell-division machinery. I will show our ability to shape live E. coli bacteria into novel shapes such as rectangles, squares, triangles and circles. I will show spatiotemporal oscillations of Min proteins – associated with cell division – in such artificial geometries of live E. coli cells [2]. (ii) Proteins and DNA in nanofabricated chambers. We use a bottom up approach to study the basic divisome components in vitro exploiting the full control provided by nanochambers. This will resolve the spatial organization of the fascinating patterns of Min proteins and chromatin that dictate the localization of the division ring. (iii) Liposomes on chip. We have developed a new chip-based technology to generate liposomes for exploring synthetic cell division [3]. We plan to use both microfluidic constrictions and a biomimetic approach (encapsulation of divisome proteins such as FtsZ) to induce liposome splitting, thus enabling a simplified form of synthetic cell division.

We believe that our mix of nanophysics and synthetic will yield insight into the biophysical underpinnings of cellular reproduction and ultimately will lead to liposomes that will be able to divide autonomously.

References: [1] Y. Caspi and C. Dekker, Systems and Synthetic Biology 8, 249-269 (2014)

[2] F. Wu, B.G.C. van Schie, J.E. Keymer, C. Dekker, Nature Nanotechnology 10, 719–726 (2015)

[3] S. Deshpande, Y. Caspi, A. Meijering, M. Jiménez, G. Rivas, C. Dekker, Nature Comm., in print

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