The KIAA1217 Knockout HT29 Polyclonal Cells comprise a CRISPR/Cas9-edited polyclonal population derived from the HT29 human colorectal adenocarcinoma cell line, designed to disrupt the KIAA1217 gene. This loss-of-function model offers a versatile system for investigating the molecular functions of KIAA1217 in colorectal cancer biology. The polyclonal configuration provides a heterogeneous collection of gene-disrupted cells, enabling robust assessment of phenotypic changes without the clonal bias inherent to single-cell-derived lines. By ablating KIAA1217 expression, researchers can interrogate its contributions to cell adhesion, migration, and cytoskeletal dynamics within a pathologically relevant epithelial background.
HT29 cells serve as a well-established model for colorectal cancer research, originating from a primary colon adenocarcinoma. These cells retain epithelial characteristics, forming polarized monolayers and expressing intestinal differentiation markers such as villin and mucins. The HT29 genetic landscape includes oncogenic mutations in APC, TP53, and BRAF V600E, which drive constitutive Wnt/??-catenin signaling and aberrant proliferation. Consequently, HT29 cells are widely employed to dissect tumor cell biology, epithelial barrier function, and therapeutic responses, providing a clinically pertinent context for studying gene function in colorectal cancer.
KIAA1217 is predicted to encode a large scaffolding protein featuring coiled-coil motifs, facilitating protein-protein interactions that tether the actin cytoskeleton to cell adhesion complexes. Within the signaling network, KIAA1217 is hypothesized to operate downstream of the ??-catenin/TCF transcription factor complex and undergo epigenetic silencing via promoter hypermethylation. It is implicated in regulating Rho family GTPases (RhoA, Rac1), focal adhesion kinase (FAK), and matrix metalloproteinases (MMP2, MMP9), ultimately influencing E-cadherin expression and adherens junction integrity. Predicted interactors include cytoskeletal components such as actin and filamin A, positioning KIAA1217 at the intersection of Wnt/??-catenin, integrin, and actin remodeling pathways.
In the HT29 background, KIAA1217 knockout enables precise dissection of its role in maintaining epithelial barrier integrity and modulating oncogenic signaling. Given the HT29 mutations in APC and BRAF, which constitutively activate Wnt and MAPK pathways, the KIAA1217 disruption can reveal how this scaffold protein integrates signals to control cell migration and invasion. This model allows researchers to assess whether KIAA1217 loss exacerbates or mitigates malignant phenotypes, such as anchorage-independent growth or metastatic potential, and to explore its involvement in cross-talk between adhesion receptors and the cytoskeleton during colorectal cancer progression.
Key applications encompass functional elucidation of KIAA1217 via transcriptomic profiling (RNA-seq), protein expression analysis (Western blotting, RT-qPCR), and cellular phenotyping using wound-healing, transwell migration, and invasion assays. Immunofluorescence staining for F-actin and adhesion markers, co-immunoprecipitation of protein complexes, and xenograft tumor growth studies further support mechanistic investigations. This knockout model is also suited for drug target validation and biomarker discovery, leveraging assays such as transepithelial electrical resistance (TEER) to monitor barrier function. For further information and customization options, please contact Ascent Research.
