The ITIH2 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population for functional studies of ITIH2 in a near-haploid fibroblast-like background. This product consists of a heterogeneous pool of HAP1 cells harboring CRISPR/Cas9-mediated disruptions of the ITIH2 locus, supplied without monoclonal selection. The polyclonal format provides a ready-to-use loss-of-function model for extracellular matrix biology and inflammation research, enabling dissection of ITIH2-dependent mechanisms without the investment of clonal isolation.
HAP1 is a near-haploid human fibroblast-like cell line derived from KBM-7 chronic myeloid leukemia cells. Its haploid genome simplifies genetic manipulation, and its adherent growth and fibroblast morphology facilitate extracellular matrix interaction and migration assays. HAP1 retains key pathways for inflammation and cancer studies, and the single-copy genome minimizes confounding wild-type allele effects, enhancing knockout phenotype interpretation. This genetic tractability and biological relevance make HAP1 an ideal host for functional genomics.
ITIH2 encodes heavy chain 2 of inter-alpha-trypsin inhibitor (I??I), which is covalently attached to hyaluronan by TSG-6 (TNFAIP6) to form stable matrices. These complexes, interacting with CD44, regulate adhesion, migration, and inflammation. ITIH2 also inhibits serine proteases like trypsin, plasmin, and neutrophil elastase via bikunin (AMBP). Transcription is regulated by IL-6, TNF-??, and glucocorticoids, linking ITIH2 to acute and chronic inflammation. Together with ITIH1, ITIH3, and ITIH4, its heavy chain is essential for hyaluronan matrix stabilization in tissue remodeling and immune cell trafficking.
In HAP1, ITIH2 knockout allows unambiguous study of hyaluronan matrix assembly and protease inhibition, avoiding the complexity of polyploid models. The near-haploid genome ensures clear loss-of-function phenotypes for secreted factors. These cells can dissect CD44 signaling and assess impacts on matrix integrity and inflammatory responses. Their fibroblastic nature suits modeling stromal ECM production, providing a robust platform for ECM-focused research.
Research applications include immunofluorescence for hyaluronan matrix, scratch wound or transwell migration/invasion assays, and protease activity measurements with chromogenic substrates. IL-6 or TNF-?? stimulation probes cytokine-dependent ITIH2 effects on matrix stabilization. The cells are applicable to psychiatric disease mechanisms, including schizophrenia and bipolar disorder via neuronal co-culture or transdifferentiation, as well as pre-eclampsia and cancer metastasis research. For further details on experimental design, contact Ascent Research.
