The KIF3B Knockout HeLa Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the human KIF3B gene in HeLa cells, creating a robust loss-of-function model for dissecting kinesin-2-dependent processes. This polyclonal pool is generated through CRISPR/Cas9-mediated gene disruption, providing a heterogeneous mixture of knockout alleles that avoids the ackgroundred drift associated with single-cell clones and enables functional studies in a bulk population. As a polyclonal knockout product, it is particularly suited for researchers investigating ciliary biology and intracellular transport without the confounding influence of clonal variation.
The host HeLa cell line, derived from a HPV18-positive cervical adenocarcinoma, is an immortalized, adherent epithelial line widely employed as a model for human cell biology. HeLa cells exhibit robust growth characteristics and are amenable to a diverse array of genetic manipulations, making them a versatile chassis for studying gene function. Under appropriate conditions, such as serum starvation, HeLa cells are capable of ciliogenesis, thereby providing a tractable system to examine cilia assembly and associated signaling pathways.
KIF3B encodes the motor subunit of the heterotrimeric kinesin-2 complex, which also includes the motor partner KIF3A and the adaptor protein KAP3 (KIFAP3). This motor complex drives anterograde intraflagellar transport (IFT) along microtubules, a process essential for ciliogenesis and the trafficking of cargo within cilia and cytoplasm. KIF3B expression is regulated transcriptionally by FOXJ1 and RFX3, and its activity can be modulated by Aurora A kinase and serum starvation. The kinesin-2 complex interacts with IFT-B proteins, including IFT88 and IFT20, as well as the dynactin component p150Glued, to transport ciliary cargo such as the Hedgehog signaling components SMO and GLI transcription factors (GLI2). Consequently, KIF3B is a critical node linking motor-driven transport to ciliary assembly and Hedgehog-responsive transcriptional outputs.
Knockout of KIF3B in HeLa cells disrupts kinesin-2 motor function, leading to defective ciliogenesis and impaired ciliary-dependent signaling, including the Hedgehog and Wnt pathways. Given the HPV-positive cervical cancer origin, this polyclonal knockout model provides a unique platform to explore the interplay between oncogenic transformation and ciliary dysfunction. The heterogeneous knockout population recapitulates the variability seen in tumor contexts and is valuable for studying how loss of KIF3B affects cellular processes such as proliferation, migration, and signaling in a cancer-relevant background.
This product is well-suited for a broad range of experimental applications. Researchers can employ immunofluorescence staining with acetylated tubulin to assess cilia formation, Western blotting to confirm KIF3B depletion, and RT-qPCR to measure expression of ciliary target genes. Live imaging of GFP-tagged IFT particles enables direct visualization of transport defects, while GLI-luciferase reporter assays quantify Hedgehog pathway activity. Additionally, wound healing migration assays can be used to investigate the role of KIF3B in cell motility. This model is also appropriate for drug screening campaigns aiming to identify modulators of ciliogenesis or ciliary signaling. For additional information, please contact Ascent Research.
