CD44 Knockout HT29 Polyclonal Cells comprise a CRISPR/Cas9-mediated polyclonal knockout cell population derived from the HT29 human colorectal adenocarcinoma cell line, providing a robust loss-of-function model for the CD44 gene in a clinically relevant epithelial context. This polyclonal product preserves population-level heterogeneity while ensuring targeted disruption of CD44, enabling detailed investigation of CD44-dependent functions without the limitations of clonal selection.
The HT29 host cell line was established from a primary colorectal adenocarcinoma of a 44-year-old Caucasian female and harbors a BRAF V600E mutation, TP53 mutation, and wild-type KRAS and APC status with proficient mismatch repair. As differentiated epithelial cells capable of forming polarized monolayers, HT29 cells are widely used to model colorectal carcinoma biology, including epithelial barrier function, differentiation programs, and oncogenic signaling networks.
CD44 encodes a transmembrane receptor for hyaluronic acid that orchestrates cell adhesion, migration, proliferation, and survival. Upon ligand binding, CD44 recruits ezrin/radixin/moesin (ERM) adaptor proteins, leading to Src kinase activation and downstream engagement of PI3K/AKT, MAPK/ERK, and NF-??B cascades. CD44 is transcriptionally regulated by NF-??B, AP-1, SP1, and the ??-catenin/TCF complex, and in turn modulates expression of MMP-9, cyclin D1, Bcl-2, and c-Myc. Interacting partners such as EGFR, c-Met, and osteopontin further embed CD44 within Rho GTPase signaling and ??-catenin stabilization networks, underscoring its role in tumor progression and therapy resistance.
In the HT29 colorectal cancer model, CD44-driven oncogenic functions intersect with the BRAF V600E?Cactivated MAPK/ERK pathway, making this knockout particularly valuable for dissecting CD44-dependent signaling in a mutant BRAF background. Disruption of CD44 is expected to impair hyaluronan-mediated adhesion and attenuate key mitogenic and anti-apoptotic signals, potentially altering invasive capacity and stemness traits. This polyclonal knockout thus enables precise evaluation of CD44 contributions to colorectal cancer biology without confounding KRAS or APC mutations.
Researchers can employ this polyclonal knockout population in a wide range of functional assays, including hyaluronic acid adhesion assays, transwell migration and invasion, wound healing, and sphere formation to interrogate metastatic and cancer stem cell phenotypes. Molecular analyses via Western blotting, RT-qPCR, phospho-AKT and phospho-ERK profiling, co-immunoprecipitation, and immunofluorescence can delineate signaling alterations. The cells are also suited for drug sensitivity screens of CD44-targeted therapeutics. For further details, please contact Ascent Research.
