Building biomimetic asymmetric lipid membranes (BEP)
Cells are physically isolated from their extracellular environment by a dynamic lipid bilayer with proteins inserted and anchored known as plasma membrane. Such bilayers are formed by many different lipids that provide them with particular biophysical properties and functionality. A well conserved property across organisms of all living kingdoms, from bacteria to animal and plant cells, is the membrane asymmetry. This means that the lipid content of each monolayer that forms the membrane is different, and this plays fundamental role in numerous cellular functions.
In our lab, we aim to build synthetic cells using a bottom-up approach, that is by reconstituting cellular elements and functions from single isolated components. Cell-sized giant unilamellar vesicles (GUVs) are the most common model architectures used in bottom-up synthetic biology to recreate a plasma membrane that encloses cell-like compartment. These GUVs are also extensively employed in membrane biophysics research. However, the model membranes employed in most of those studies usually lack an asymmetric lipid distribution.
In this project you will produce GUVs with varying lipid compositions for each monolayer with the aim to build biomimetic asymmetric lipid membranes. For this purpose, you will use a method developed in our group called emulsion droplet interface crossing encapsulation (eDICE). Then, you will employ confocal fluorescence microscopy to assess the distribution of different lipids within the membrane.