KIF3B Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the near-haploid human HAP1 cell line. This product provides a heterogeneous pool of cells carrying targeted disruptions in the KIF3B gene, resulting in loss of KIF3B protein expression. As a polyclonal knockout population, it retains the inherent advantages of rapid experimental deployment without the need for single-cell cloning, making it ideal for functional genomics and high-throughput screening applications.
The parental HAP1 cell line originates from the chronic myelogenous leukemia (CML)-derived KBM-7 line and exhibits a near-haploid karyotype with adherent, fibroblast-like morphology. Its haploid nature simplifies knockout generation and phenotypic interpretation, as only one allele typically requires editing to achieve functional gene disruption. HAP1 cells are widely employed in CRISPR-based functional screens and gene-editing studies due to their stable growth characteristics and ease of manipulation.
KIF3B encodes a subunit of the heterotrimeric kinesin-2 motor complex with KIF3A and KAP3 (KIFAP3). This motor drives anterograde intraflagellar transport (IFT), moving IFT particles and ciliary membrane proteins along axonemal microtubules from the ciliary base to the tip, making it essential for ciliogenesis. KIF3B knockout abrogates anterograde IFT, causing defective ciliary assembly and impaired Hedgehog signaling, as pathway components SMO, GLI2, and SUFU become mislocalized. The kinesin-2 complex interacts with IFT-B members IFT88 and IFT172, as well as tubulin and dynein motors. KIF3B expression is regulated by serum deprivation, cell cycle arrest, and RFX transcription factors, placing it downstream of these cues in ciliary trafficking and signaling.
In the HAP1 background, KIF3B knockout provides a powerful model to dissect cilia-dependent signaling pathways with minimal genetic redundancy. The loss of KIF3B disrupts Hedgehog signal transduction, a pathway implicated in development and oncogenesis. This model is particularly relevant for studying ciliopathies such as retinitis pigmentosa and polycystic kidney disease, as well as KIF3B-associated cancers including breast and colorectal carcinomas. The combination of CRISPR-mediated gene disruption and the HAP1 cell line??s haploid genetics facilitates clear interpretation of genotype-phenotype relationships in ciliary biology.
This polyclonal knockout population enables diverse experimental applications. Immunofluorescence microscopy with ciliary markers (acetylated tubulin, Arl13b) reports on cilia morphology, while Western blotting detects KIF3B, IFT proteins, and GLI processing. RT-qPCR quantifies Hedgehog target genes such as GLI1 and PTCH1. Co-immunoprecipitation probes KIF3B interactors; migration/invasion assays assess cancer-relevant phenotypes; and flow cytometry measures cilia length or cell cycle distribution. These cells support functional studies of intraflagellar transport, ciliogenesis, and Hedgehog signaling in basic research and drug discovery. For further details, contact Ascent Research.
