Optimization of a deep mutational scanning workflow to improve quantification of mutation effects on protein-protein interactions.

ABSTRACT: Deep Mutational Scanning (DMS) assays are powerful tools to study sequence-function relationships by measuring the effects of thousands of sequence variants on protein function. During a DMS experiment, several technical artefacts might distort non-linearly the functional score obtained, potentially biasing the interpretation of the results. We therefore tested several technical parameters in the deepPCA workflow, a DMS assay for protein-protein interactions, in order to identify technical sources of non-linearities. We found that parameters common to many DMS assays such as amount of transformed DNA, timepoint of harvest and library composition can cause non-linearities in the data. Designing experiments in a way to minimize these non-linear effects will improve the quantification and int

SUBMITTER: Bendel AM 

PROVIDER: S-EPMC11194945 | biostudies-literature | 2024 Jun

REPOSITORIES: biostudies-literature