The KIF3B Knockout HT29 Polyclonal Cells consist of a CRISPR/Cas9-edited polyclonal HT29 colorectal adenocarcinoma cell population with targeted disruption of KIF3B. This loss-of-function model avoids single-cell cloning artifacts by using a heterogeneous pool of knockout cells generated via CRISPR/Cas9-mediated gene disruption. It enables robust functional analyses of KIF3B in signaling and trafficking pathways.
The parental HT29 cell line is a widely used human colorectal adenocarcinoma line established from a primary tumor, and it serves as an important in vitro model of intestinal epithelial biology and colorectal cancer. These adherent cells retain key characteristics of colorectal tumors, including aberrant Wnt pathway activation and the capacity for differentiation under defined conditions. HT29 cells are extensively utilized to study colorectal cancer progression, drug responses, and mechanisms of metastasis. Their inherent properties make them a suitable host for investigating genes implicated in colorectal tumorigenesis and ciliary signaling.
KIF3B is a core subunit of the heterotrimeric kinesin-2 motor, partnering with KIF3A and KAP3 to transport cargo anterogradely along microtubules. This motor is essential for intraflagellar transport and primary cilium assembly, where it mediates Hedgehog signaling by trafficking Smoothened (SMO) and enabling activation of GLI transcription factors. KIF3B interacts with IFT88 and the IFT-B complex, and its expression is controlled by FOXJ1 and RFX transcription factors. The motor also associates with the chromosomal passenger complex, contributing to mitotic spindle organization. Thus, KIF3B bridges ciliary homeostasis, signal transduction, and cell division.
In HT29 cells, KIF3B disruption likely impairs ciliogenesis and attenuates Hedgehog and Wnt pathway crosstalk, potentially reducing GLI-driven transcription. Loss of KIF3B-dependent transport may also hamper cell migration and invasion, processes key to colorectal cancer metastasis. This polyclonal knockout model thus provides a valuable platform for dissecting KIF3B’s roles in colorectal adenocarcinoma progression and cilia-dependent signaling, offering insights into tumor cell motility and drug sensitivity.
Research applications include colorectal cancer metastasis, cilia biology, and intracellular trafficking studies. Typical assays encompass immunofluorescence for ciliary markers (acetylated tubulin, Arl13b), Western blotting, Boyden chamber migration/invasion assays, MTT proliferation assays, RNA-seq, and flow cytometry for cell cycle analysis. For additional information, please contact Ascent Research.
