The CCM2 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from the HT29 human colorectal adenocarcinoma cell line. This product provides a loss-of-function model through CRISPR/Cas9-mediated disruption of the endogenous CCM2 gene. The polyclonal pool retains the heterogeneous genetic background of the bulk population, enabling broad screening without clonal selection bias.
HT29 cells are an epithelial colon adenocarcinoma model capable of forming polarized monolayers, widely used to study intestinal barrier function and colorectal cancer. They express key junctional proteins and harbor a BRAF V600E mutation, making them a relevant host for investigating scaffolding proteins in epithelial integrity.
CCM2 (malcavernin) is a scaffold protein that bridges CCM1 (KRIT1) and CCM3 (PDCD10) in the CCM complex. It negatively regulates RhoA-ROCK signaling to control actin cytoskeleton dynamics and barrier integrity, and suppresses the MEKK3-MKK5-ERK5 pathway by direct interaction with MEKK3. The complex integrates signals from integrin-mediated adhesion and shear stress, with upstream regulators such as integrin ??1 and Rap1 modulating its assembly. Downstream effectors include RhoA GTPase, ERK5 kinase, VE-cadherin-based junctions, and the actin cytoskeleton. Interactions with ICAP1 and SMURF1 further fine-tune complex stability and localization. Disruption of CCM2 unleashes sustained RhoA-ROCK activation and ERK5 signaling, promoting stress fiber formation and junctional breakdown.
Loss of CCM2 in HT29 cells provides an epithelial model to study barrier function and cancer cell adhesion. These cells form tight junctions and polarized architectures, allowing measurement of trans-epithelial electrical resistance and localization of junctional proteins. CCM2 ablation is expected to compromise cell?Ccell adhesion, increase stress fibers, and alter RhoA-mediated contractility, paralleling defects observed in vascular malformations in an intestinal epithelial context.
These polyclonal knockout cells are suited for RhoA GTPase activity assays, Western blotting for phosphorylated ERK5, TEER analysis, immunofluorescence for VE-cadherin/ZO-1, and wound healing migration assays. RNA sequencing and apoptosis or proliferation assays further support drug screening and mechanistic studies targeting colorectal cancer and vascular malformation pathways. For further information or assistance, contact Ascent Research.
