The DSE Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population of HAP1 cells, disrupting the DSE gene to eliminate dermatan sulfate epimerase function. This non-clonal pool provides a genetically diverse loss-of-function model for studying glycosaminoglycan biology and extracellular matrix (ECM) regulation.
HAP1 is a near-haploid human fibroblast-like cell line derived from the KBM-7 chronic myeloid leukemia line. Its near-haploid genome facilitates efficient gene disruption and functional genomics, and the cells retain mesenchymal features relevant for ECM and glycosaminoglycan studies.
DSE encodes dermatan sulfate epimerase, which catalyzes the conversion of glucuronic acid to iduronic acid within nascent dermatan sulfate chains, a key modification that introduces conformational flexibility and enhances binding to diverse growth factors and matrix proteins. This reaction is critical for generating the structural diversity of glycosaminoglycans and is tightly regulated by signals such as TGFB1 and inflammatory cytokines. DSE functions in a biosynthetic complex with DSEL, sulfotransferases CHST14 and D4ST1, and other glycosaminoglycan synthases. Its epimerase activity directly shapes the sulfation patterns of dermatan sulfate proteoglycans, including decorin and biglycan, ultimately influencing downstream processes such as FGF2 and HGF binding, as well as collagen fibrillogenesis. Thus, DSE is a central node linking glycosaminoglycan structure to ECM organization and growth factor signaling.
Knocking out DSE in HAP1 cells abrogates iduronic acid formation, yielding structurally altered dermatan sulfate that compromises ECM integrity and growth factor signaling. This phenotype mirrors connective tissue defects seen in Ehlers-Danlos syndrome musculocontractural type 2 and is pertinent to fibrosis and cancer metastasis research. The polyclonal nature avoids clonal bias while the haploid background ensures efficient knockout.
Researchers can exploit the DSE knockout polyclonal pool in a broad range of experimental workflows. Confirmation of gene disruption and loss of epimerase activity can be achieved via western blotting for DSE protein and HPLC/MS analysis of dermatan sulfate disaccharide composition. Changes in proteoglycan expression and distribution are detectable by immunofluorescence and RT-qPCR for decorin and biglycan. To assess functional consequences, cell migration and invasion assays can model cancer metastatic behavior, while FGF2 and HGF binding assays quantify altered growth factor interactions. Collagen gel contraction assays provide a readout of ECM contractility, relevant to fibrosis and connective tissue disorders. Moreover, this model can be employed in drug screens targeting glycosaminoglycan biosynthesis or ECM remodeling pathways. For further information, please contact Ascent Research.
