Optimization of a deep mutational scanning workflow to improve quantification of mutation effects on protein-protein interactions
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...
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Veröffentlicht in: | BMC genomics 2024-06, Vol.25 (1), p.630-16, Article 630 |
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Sprache: | eng |
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Zusammenfassung: | 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 interpretation of mutation effects. |
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ISSN: | 1471-2164 1471-2164 |
DOI: | 10.1186/s12864-024-10524-7 |