The IQGAP1 Knockout HT29 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal population of HT29 colorectal adenocarcinoma cells harboring targeted disruption of the IQGAP1 gene. This loss-of-function model enables systematic investigation of IQGAP1 scaffolding functions in epithelial cell biology and oncogenic signaling.
HT29 is a well-characterized human colon cancer cell line with epithelial morphology, originally derived from a primary colon carcinoma. It carries an activating BRAF V600E mutation, along with inactivating mutations in APC and SMAD4, while retaining wild-type p53. These genetic features make HT29 a relevant model for studying colorectal cancer with constitutive Wnt pathway activation and altered TGF-??/BMP signaling.
IQGAP1 is a multi-domain scaffold protein that integrates signals from Rho GTPases (Cdc42, Rac1), Ca2+/calmodulin, and growth factor receptors to regulate cell adhesion, migration, and proliferation. It directly binds ??-catenin, E-cadherin, actin, calmodulin, MEK, ERK, and APC, thereby bridging adherens junction stability with actin cytoskeleton dynamics. IQGAP1 acts downstream of EGF and integrin signaling, promoting ??-catenin nuclear translocation, E-cadherin stabilization, actin polymerization, and ERK pathway activation. In the Wnt signaling context, IQGAP1 scaffolds ??-catenin/E-cadherin complexes at the membrane and modulates ??-catenin transcriptional activity, while in MAPK/ERK signaling it facilitates MEK-ERK complex formation.
In HT29 cells, APC mutations lead to aberrant ??-catenin accumulation, yet IQGAP1 still modulates the balance between ??-catenin adhesive and transcriptional pools. IQGAP1 knockout in this background disrupts ??-catenin/E-cadherin interactions, weakening cell-cell adhesion and likely altering ??-catenin nuclear signaling. Concurrently, attenuated IQGAP1-dependent ERK activation reduces migratory and invasive capacity. The polyclonal nature of the knockout population guards against clonal artifacts, offering a robust platform for investigating colorectal cancer metastasis, E-cadherin dynamics, and crosstalk between Wnt and MAPK pathways.
This knockout product is suited for diverse experimental applications, including transwell migration and invasion assays, immunofluorescence analysis of actin stress fibers and adherens junctions, western blotting for ??-catenin/E-cadherin and phospho-ERK, co-immunoprecipitation of ??-catenin complexes, and proliferation assays. It supports research into colorectal cancer metastasis, drug resistance mechanisms, and tumor microenvironment interactions. For further information, please contact Ascent Research.
