biostudies-literatureUnknown44Norouzi MWhen found on the cell surface, Vimentin, a cytoskeletal protein, engages in multiple pathophysiological processes. Here, in human fibroblasts and endothelial cells, we used CRISPR-Cas9 to insert a hemagglutinin tag at six different loci within the <i>vim</i> gene. We then utilized tag immunodetection to define cell surface Vimentin's (csVim) membrane topology, followed by proximity biotinylation to identify its extracellular interactome; validating a subset of hits with a quantitative in vitro binding assay.Biochemistry and biophysics reports102242https://www.ebi.ac.uk/biostudies/studies/S-EPMC12447900biostudies-literatureAn adaptable genomic-proteomic approach to characterize cell-surface Vimentin's membrane topology and interactome.PMC12447900Norouzi MMcCulloch CAfalseAn adaptable genomic-proteomic approach to characterize cell-surface Vimentin's membrane topology and interactome.When found on the cell surface, Vimentin, a cytoskeletal protein, engages in multiple pathophysiological processes. Here, in human fibroblasts and endothelial cells, we used CRISPR-Cas9 to insert a hemagglutinin tag at six different loci within the <i>vim</i> gene. We then utilized tag immunodetection to define cell surface Vimentin's (csVim) membrane topology, followed by proximity biotinylation to identify its extracellular interactome; validating a subset of hits with a quantitative in vitro binding assay.2025-01-01T00:00:00Z2025 Dec2026-06-03T17:17:42.302Z2026-04-29T03:12:48.204ZS-EPMC124479004097820410.1016/j.bbrep.2025.102242