IFT46 Knockout HAP1 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population derived from the near-haploid HAP1 human cell line, engineered to disrupt expression of the IFT46 gene. This product provides a heterogeneous pool of edited cells, enabling functional studies of IFT46 loss within a genetically uniform host background. The knockout introduction was performed using CRISPR/Cas9-mediated gene disruption targeting IFT46, generating a versatile model for investigating the molecular consequences of impaired intraflagellar transport (IFT) and ciliary signaling. As a polyclonal population, it preserves the natural variability of editing outcomes while maintaining the key advantages of the HAP1 system for genetic and pharmacological screens.
HAP1 cells are a male-derived adherent cell line originating from the KBM-7 chronic myeloid leukemia line, characterized by a near-haploid karyotype and fibroblast-like morphology. Their haploid nature facilitates straightforward genetic manipulation and phenotype?Cgenotype correlation, making them exceptionally suited for loss-of-function screens and targeted gene knockout studies. The HAP1 model has been widely adopted in functional genomics, cancer biology, and signal transduction research due to its stable growth properties and compatibility with high-content imaging and biochemical assays. The near-haploid genome simplifies interpretation of knockout phenotypes, as recessive mutations are directly expressed without confounding diploid compensation, thereby enhancing the clarity of experimental readouts.
IFT46 is a core subunit of the intraflagellar transport complex B (IFT-B), which is essential for anterograde trafficking of ciliary components along the axoneme. It is transcriptionally regulated by RFX transcription factors and FOXJ1 and directly interacts with other IFT-B components such as IFT20, IFT52, and IFT88, as well as the kinesin-2 motor subunit KIF3B, to drive active transport toward the ciliary tip. Disruption of IFT46 function prevents the ciliary accumulation of key signaling receptors including SMO, PTCH1, and PDGFR??, thereby attenuating Hedgehog pathway activation downstream of GLI transcription factors. Additionally, IFT46 deficiency impairs Wnt/??-catenin signaling by altering ??-catenin stabilization, highlighting its integrative role in cilia-dependent signal transduction networks. This positions IFT46 as a critical node linking ciliary structural integrity to developmental and oncogenic signaling cascades.
In the HAP1 haploid background, IFT46 knockout provides a genetically clean platform for dissecting ciliogenesis and ciliary signaling. The loss of IFT46 abrogates anterograde IFT and consequently blocks primary cilium assembly, a process readily visualized by immunofluorescence for ciliary markers such as acetylated tubulin and ARL13B. This defect leads to profound suppression of Hedgehog transcriptional output, as GLI1 expression and GLI-luciferase reporter activity are diminished, and also perturbs PDGFR?? signaling. The model therefore serves as a valuable tool for studying the molecular pathology of ciliopathies??including skeletal dysplasias, retinitis pigmentosa, and polycystic kidney disease??and for exploring Hedgehog-driven cancers such as basal cell carcinoma and medulloblastoma, where constitutive pathway activation is a hallmark.
Researchers can employ this knockout model for a variety of advanced applications. Ciliogenesis assays, when combined with quantitative immunofluorescence, allow systematic evaluation of ciliary assembly and receptor trafficking defects. Hedgehog reporter assays (Gli-luciferase), RT-qPCR profiling of GLI1 and PTCH1, and Western blot analysis of IFT46 expression provide robust readouts of pathway activity and knockout efficiency. The cells are also amenable to high-throughput screening of small-molecule Hedgehog inhibitors or modulators of ciliary trafficking. These applications make the IFT46 Knockout HAP1 Polyclonal Cells a versatile resource for functional genomics, drug discovery, and mechanistic studies of cilia-dependent signaling. For further technical details or support, please contact Ascent Research.
