The IFT20 Knockout HAP1 Polyclonal Cells are a polyclonal knockout cell population generated by CRISPR/Cas9-mediated disruption of the IFT20 gene in the human near-haploid HAP1 cell line. This product provides a genetically defined loss-of-function model for investigating the roles of intraflagellar transport protein 20 (IFT20), a critical subunit of the IFT-B complex. The polyclonal format ensures a heterogeneous knockout population suitable for robust functional studies and pooled screening applications.
The HAP1 cell line, derived from the chronic myeloid leukemia KBM-7 line, exhibits adherent fibroblastoid morphology and a near-haploid karyotype. Its haploid genetic content makes it an attractive model for genetic screening and knockout analysis, enabling clean genotype-phenotype correlations. HAP1 cells have been widely adopted for CRISPR-based functional genomics and retain the capacity to form primary cilia under appropriate conditions, providing a valuable platform for ciliary biology research.
IFT20 encodes a core component of the IFT-B anterograde trafficking complex, essential for ciliogenesis and ciliary cargo transport. IFT20 bridges the IFT-B particle with Golgi-associated proteins golgin-160 and GMAP210, facilitating targeted delivery of ciliary membrane proteins such as SMO and polycystins. Acting downstream of ciliogenic transcription factors FOXJ1 and RFX, IFT20 is indispensable for primary cilium assembly and Hedgehog signal transduction. SHH binding to PTCH1 relieves inhibition of SMO, which enters the cilium via IFT-dependent trafficking, leading to GLI transcription factor activation. IFT20 knockout ablates anterograde transport, blocking SMO entry and GLI activation, and also disrupts Wnt/planar cell polarity signaling dependent on proper ciliary positioning.
In the ciliated HAP1 background, IFT20 loss-of-function produces robust ciliogenesis defects, recapitulating core features of ciliopathies including polycystic kidney disease, retinal degeneration, and skeletal dysplasias. The near-haploid genome enhances knockout phenotype penetrance, providing a clean system to study primary cilium assembly and signaling. The leukemic origin of HAP1 cells offers an opportunity to explore links between ciliary signaling and hematopoietic malignancies. Researchers can exploit this model to dissect the roles of ciliary Hedgehog and Wnt signaling in proliferation, differentiation, and drug response, leveraging the cell line’s genetic simplicity.
Representative applications include immunofluorescence staining for ciliary markers (ARL13B, acetylated tubulin) to quantify cilia morphology, western blotting for IFT-B subunits and GLI factors, and qPCR analysis of Hedgehog target genes (PTCH1, GLI1). The model supports high-content screening for ciliogenic compounds, wound-healing assays for planar polarity, and paracrine signaling studies. The polyclonal knockout format is ideal for pooled CRISPR screens and dose-response studies in drug discovery. For further information, contact Ascent Research.
