The KIF3B Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the widely used HEK293T cell line. This product provides a genetically diverse pool with targeted KIF3B gene disruptions, enabling loss-of-function studies without clonal selection. The polyclonal format offers a robust model minimizing clonal artifacts from single-cell-derived knockouts. The CRISPR/Cas9-mediated gene disruption introduces stable modifications in the KIF3B locus, yielding a mixed population that reflects functional consequences across diverse genetic backgrounds.
The host cell line, HEK293T, is a human embryonic kidney epithelial line stably expressing SV40 large T antigen, renowned for high transfection efficiency, rapid growth, and robust protein expression. It serves as a cornerstone for recombinant protein production and lentiviral packaging. This background provides a relevant context for kidney-derived epithelial signaling studies and its manipulative ease facilitates detailed mechanistic dissection. Under serum starvation, HEK293T cells can form primary cilia, enabling ciliogenesis-focused research.
KIF3B encodes a crucial subunit of the heterotrimeric kinesin-2 motor complex, which also includes KIF3A and the kinesin-associated protein KAP3 (KIFAP3). This motor complex drives anterograde intraflagellar transport (IFT) along ciliary axonemes, a process essential for the assembly, maintenance, and signaling functions of primary cilia. KIF3B interacts with IFT particles and adaptors to transport ciliary membrane components and Hedgehog transducer Smoothened. KIF3B acts downstream of cargo adaptor proteins and IFT complex assembly factors; its activity is integral to Hedgehog and Wnt signaling by enabling proper GLI transcription factor localization and activation. Disruption of KIF3B therefore impairs IFT, leading to defective cilia formation and attenuated ciliary-based signal transduction.
In the HEK293T context, KIF3B knockout provides a powerful tool to dissect the molecular requirements for ciliogenesis and ciliary signaling in epithelial cells. KIF3B loss compromises primary cilia assembly and ciliary receptor trafficking, rendering the model valuable for studying ciliopathies such as polycystic kidney disease, retinitis pigmentosa, and Bardet-Biedl syndrome. The HEK293T background also permits efficient complementation experiments, enabling structure-function analysis of the kinesin-2 complex. The polyclonal population mirrors patient genetic heterogeneity, offering a relevant drug screening platform.
Researchers can employ the KIF3B Knockout HEK293T Polyclonal Cells in applications including Western blotting for knockout confirmation, immunofluorescence to assess cilia morphology via acetylated tubulin or Arl13b staining, and RT-qPCR to measure Hedgehog target gene expression (e.g., GLI1, PTCH1). Functional assays such as flow cytometry for cell cycle analysis or cell migration can reveal phenotypic consequences. These cells are ideal for high-content screening of ciliogenesis modulators or Hedgehog restorers. For more information, contact Ascent Research.
