Design of a mutational library to measure the stability of all possible mutations in sod1

Identifier:  TFG:6614

Handle: https://hdl.handle.net/20.500.11797/TFG6614

Authors:  Quiroga, Tomás

Abstract:
Superoxide Dismutase 1 (SOD1) is a protein that neutralizes Reactive Oxygen Species (ROS). Mutations in SOD1 can cause ALS. This study aims to identify mutations that lead to protein misfolding using Deep Mutational Scanning (DMS). The DMS approach involves creating a mutational library of SOD1, using a high-throughput protein abundance assay (PCA) to select it, and deep sequencing to measure the impact of mutations. By fusing SOD1 with the DHFR enzyme, we conducted an abundance assay to quantify the effect of mutations. We created a library of 6000 distinct SOD1 mutations to test their impact on protein folding and plan to use the results to distinguish between pathogenic and non-pathogenic mutations.