While de novo protein engineering of soluble proteins has advanced significantly, designing complex multi-pass transmembrane proteins remains difficult. The goal of this project is to develop a framework that maps how specific sequences control a transmembrane protein’s localization, orientation, and cellular impact. We will introduce libraries of de novo and native transmembrane constructs into cells and analyze them using a combination of high-throughput techniques. Multiplexed Error Robust Fluorescence In Situ Hybridization(MERFISH) will be used to identify constructs and measure endoplasmic reticulum stress, while oligo-conjugated immunofluorescence and split fluorescent assays will physically map the protein’s location and topology. Ultimately, this comprehensive dataset will be used to train a predictive model to improve the future design of these complex proteins.

This project has allowed me to explore my interest in experimental design, imaging, and computation, and how these approaches can work together to answer questions in cell biology. Through SuperUROP, I am very excited to continue applying these approaches to fundamental questions about membrane protein localization and trafficking.