PhD: Role of the cytoskeleton in cell membrane electroporation and cargo translocation

New genome editing tools promise to repair cell functions at the source. However, they require the delivery of nucleic acids and/or gene editing enzymes directly into cells. Electroporation by transient disruption of the membrane under electric pulses can in principle deliver cargo molecules straight to the cytosol. However, the mechanisms that determine the efficiency of electroporation across the cell membrane are unknown. We recently discovered that the actin cytoskeleton, a crosslinked network of filamentous proteins that lines the cell membrane, plays an important role in the barrier function of the cell surface. The aim of this experimental PhD project is to reveal the biophysical mechanisms by which the actin cortex and the membrane together govern electro-transfer of nucleic acids and proteins across the cell membrane. To this end, you will harness techniques developed by the Koenderink lab to create synthetic cell-like lipid vesicles with tunable cytoskeleton-membrane interactions. To measure the mechanical behavior of the membrane-cytoskeleton interface, you will develop novel microfluidic aspiration devices. Finally, you will use high resolution microscopy combined with microfluidic electroporation devices to visualize the electro-transfer of model cargoes (DNA/proteins) and CRISPR-Cas gene editing molecules across the membrane of the synthetic cells. By teaming up with a PhD student in Pouyan Boukany’s lab who will study electroporation in living cells and with the team of Lea Rems for membrane simulations, we will be able to understand the mechanisms that govern electroporation efficiency as a prerequisite toward the future rational design of robust and safe methods for non-viral delivery of gene-editing molecules.

We offer an inspiring, supportive and collegial environment. The Koenderink lab is an experimental biophysics lab studying the mechanobiology of cells and tissues. We combine concepts and techniques from soft matter physics, biophysics, synthetic biology, protein engineering, and cell biology. The Koenderink lab is embedded in the TU Delft Department of Bionanoscience, which focuses on the fundamental understanding of biological processes from molecule to cell. The department features an inspiring, international environment with access to state-of-the art facilities for nanofabrication, a microscopy facility, molecular/cell biology, biochemistry, and high-performance computing for image processing.