| Literature DB >> 35524121 |
Alexandre K Dubé1,2,3,4,5, Rohan Dandage6,7,8,9,10, Soham Dibyachintan6,7,8,10,11, Ugo Dionne7,8,9,12,13, Philippe C Després6,7,8,9, Christian R Landry14,15,16,17,18.
Abstract
Deep mutational scanning (DMS) generates mutants of a protein of interest in a comprehensive manner. CRISPR-Cas9 technology enables large-scale genome editing with high efficiency. Using both DMS and CRISPR-Cas9 therefore allows us to investigate the effects of thousands of mutations inserted directly in the genome. Combined with protein-fragment complementation assay (PCA), which enables the quantitative measurement of protein-protein interactions (PPIs) in vivo, these methods allow for the systematic assessment of the effects of mutations on PPIs in living cells. Here, we describe a method leveraging DMS, CRISPR-Cas9, and PCA to study the effect of point mutations on PPIs mediated by protein domains in yeast.Entities:
Keywords: CRISPR-Cas9; Deep mutagenesis scanning; Protein-fragment complementation assay; Protein–protein interaction; Yeast
Mesh：
Year: 2022 PMID： 35524121 DOI： 10.1007/978-1-0716-2257-5_14
Source DB: PubMed Journal: Methods Mol Biol ISSN： 1064-3745