This product is a CRISPR/Cas9-edited polyclonal knockout cell population of HT29 human colorectal adenocarcinoma cells with disrupted KIF3A expression. The knockout model provides a loss-of-function tool for studying kinesin-2-mediated transport in oncogenic signaling. Its polyclonal nature avoids clonal selection artifacts, enabling robust population-level analysis of KIF3A-dependent cellular processes.
The HT29 parental line, derived from a primary colorectal adenocarcinoma in a 44-year-old female, displays adherent epithelial morphology with wild-type p53 and microsatellite stability. It serves as a well-established model for intestinal epithelial biology, capable of enterocytic differentiation and mucus production, making it particularly relevant for differentiation and barrier function studies in colon cancer research.
KIF3A encodes the motor subunit of the kinesin-2 complex, powering anterograde transport along microtubules and essential for intraflagellar transport and ciliogenesis. It traffics key signaling molecules including ??-catenin and Gli transcription factors, thereby mediating Wnt and Hedgehog pathway activation. KIF3A forms complexes with KIF3B and KAP3 (KIFAP3), interacting with APC and ??-catenin. Knockout disrupts transport, reducing ??-catenin nuclear accumulation and transcription of target genes such as MYC, CCND1, and GLI1, with upstream regulation by Wnt ligands and Sonic hedgehog and crosstalk with mTOR signaling.
In HT29 cells, KIF3A loss impairs Wnt and Hedgehog signaling pathways often hyperactivated in colorectal cancer. Disruption of ciliary and intracellular transport attenuates ??-catenin?CTCF/LEF and GLI-dependent transcription, altering proliferation, migration, and differentiation. This model thus recapitulates the dependence of colorectal adenocarcinoma cells on kinesin-2 motor function for oncogenic phenotypes and enables evaluation of KIF3A as a therapeutic target.
Applications include Western blotting for KIF3A, ??-catenin, and Gli1; RT-qPCR for MYC, CCND1, and GLI1; immunofluorescence for ciliary morphology using acetylated tubulin; wound healing and transwell invasion assays; Wnt/??-catenin (TOP/FOP) and Hedgehog luciferase reporter assays; co-immunoprecipitation of KIF3A?CKAP3 complexes; and cell viability or drug sensitivity screens. This polyclonal knockout population supports mechanistic studies, functional genomics, and drug discovery research in colorectal cancer. For additional technical details or a quotation, contact Ascent Research.
