biostudies-literatureJia YChina Scholarship CouncilARCCentre Franco-Indian pour la Promotion de la recherche AvancéeLigue Régionale contre le CancerFondation pour la Recherche Médicale200https://www.ebi.ac.uk/biostudies/studies/S-EPMC9818449biostudies-literatureUnknown12(1)Biological pathways rely on the formation of intricate protein interaction networks called interactomes. Getting a comprehensive map of interactomes implies the development of tools that allow one to capture transient and low-affinity protein-protein interactions (PPIs) in live conditions. Here we presented an experimental strategy: the Cell-PCA (cell-based protein complementation assay), which was based on bimolecular fluorescence complementation (BiFC) for ORFeome-wide screening of proteins that interact with different bait proteins in the same live cell context, by combining high-throughput sequencing method. The specificity and sensitivity of the Cell-PCA was established by using a wild-type and a single-amino-acid-mutated HOXA9 protein, and the approach was subsequently applied to sevCellsA Live Cell Protein Complementation Assay for ORFeome-Wide Probing of Human HOX Interactomes.PMC98184495503-PPJA20191209567201708070003160896119030Merabet SKundlacz CVanacker JMReboulet JHughes SHajj Sleiman NJia YTribollet VGillet BForcet CBleicher FfalseA Live Cell Protein Complementation Assay for ORFeome-Wide Probing of Human HOX Interactomes.Biological pathways rely on the formation of intricate protein interaction networks called interactomes. Getting a comprehensive map of interactomes implies the development of tools that allow one to capture transient and low-affinity protein-protein interactions (PPIs) in live conditions. Here we presented an experimental strategy: the Cell-PCA (cell-based protein complementation assay), which was based on bimolecular fluorescence complementation (BiFC) for ORFeome-wide screening of proteins that interact with different bait proteins in the same live cell context, by combining high-throughput sequencing method. The specificity and sensitivity of the Cell-PCA was established by using a wild-type and a single-amino-acid-mutated HOXA9 protein, and the approach was subsequently applied to sev2023-01-01T00:00:00Z2023 Jan2025-04-26T06:03:11.726Z2025-04-06T11:43:38.491ZS-EPMC98184493661199310.3390/cells12010200