The ITIH2 Knockout HT29 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population designed for the targeted disruption of the ITIH2 gene in the human HT29 colorectal adenocarcinoma cell line. This polyclonal pool offers a heterogeneous loss-of-function model, enabling robust analysis of ITIH2-dependent processes without clonal selection artifacts. The CRISPR/Cas9-mediated gene disruption provides a versatile platform for studying the functions of ITIH2 within the context of colorectal cancer biology.
The HT29 cell line, derived from a primary human colorectal adenocarcinoma, is a widely utilized epithelial model for colorectal cancer research. These cells exhibit an epithelial morphology and can be induced to differentiate into enterocyte-like cells under appropriate culture conditions, making them suitable for investigating drug transport mechanisms and intestinal epithelial biology. Their well-characterized genetic and phenotypic properties facilitate reliable experimental reproducibility in cancer and pharmacological studies.
ITIH2 encodes the heavy chain of inter-alpha-trypsin inhibitor (ITI) complexes, which are essential for extracellular matrix (ECM) stabilization through covalent linkage to hyaluronan (HA) mediated by TSG-6. This gene product interacts with bikunin (encoded by AMBP) and other ITI heavy chains (ITIH1, ITIH3, ITIH4) to form complexes that modulate inflammatory responses and ECM organization. Upstream regulators include the pro-inflammatory cytokines IL-6 and TNF-alpha, which activate NF-kB signaling to transcriptionally regulate ITIH2 expression. Downstream, ITIH2-containing complexes influence hyaluronan-CD44 signaling, a pathway implicated in cell adhesion, migration, and invasion. ITIH2 also acts in concert with hyaluronic acid synthase 2 (HAS2) and TSG-6 to maintain pericellular matrix architecture, thereby affecting cellular behavior and tissue homeostasis.
In HT29 colorectal cancer cells, disruption of ITIH2 is anticipated to compromise hyaluronan matrix integrity, potentially altering cell adhesion, migration, and inflammatory responses associated with tumor progression. Given the role of ITIH2 in ECM remodeling and serine protease inhibition, its knockout provides a valuable model for dissecting the molecular interplay between the ECM and tumor-associated inflammation in colorectal cancer. This system may reveal how ITIH2 loss impacts tumor microenvironment dynamics, including cytokine profiles and chemosensitivity, thereby offering insights into metastasis and therapeutic resistance.
Researchers can employ this polyclonal knockout cell pool for a wide range of functional investigations. Representative applications include tumor microenvironment studies, hyaluronan biology, ECM remodeling, and inflammation-associated colorectal cancer research. Common assays include Western blotting and RT-qPCR to confirm ITIH2 disruption, particle exclusion assays to visualize hyaluronan matrices, Boyden chamber invasion/migration tests, MTT or CCK-8 proliferation assays, cytokine profiling via ELISA arrays, and immunofluorescence staining for hyaluronan. This model is also suitable for chemosensitivity screens using agents such as 5-fluorouracil or oxaliplatin, enabling the assessment of drug response in an ITIH2-deficient background. For additional technical details or custom inquiries, please contact Ascent Research.
