EAPSI: A Bottom-up Approach to Engineering an Artificial Cell

Riche Carson, Los Angeles CA

Investigators

Abstract

Intrinsic cellular function is challenging to study due to many interconnected pathways. There are two ways to attack this problem. One can work with a fully functioning cell and attempt to turn off all the undesired signaling pathways to only leave ?on? the function of interest. This is extremely challenging. Conversely, one can address this problem using a bottom-up approach. This project will be using the latter method. Starting with the simplest of building blocks, lipid molecules, and adding proteins and internal structure the complexity will be increased. In this technique, only the cell mimicking molecules of interest need to be added. This research will be conducted in collaboration with Dr. Tae-Joon Jeon at Inha University in Incheon, South Korea. The project will leverage the state-of-the-art electrophysiology equipment and protein characterization in his laboratory. This award will support the study the self-assembly of proteovesicles, bilayer membranes containing integral membrane proteins. Using in vitro transcription translation the project will incorporate aquaporin channels into phospholipid membranes. These proteins span the bilayer and transport water molecules between the internal and external cellular environment. Their function can be probed by electrophysiology measurements that quantify the electrical conductance across individual channels. The project will compare a new, simplified method of incorporating these proteins to existing protein incorporation methods. The validation of the new technique will be an important step in convincing the community that the new technique is superior to antiquated methods. This award is funded in collaboration with the National Research Foundation of Korea.