The IFT172 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal cell population derived from the near-haploid HAP1 line, engineered for loss-of-function analysis of intraflagellar transport and ciliary signaling. These cells carry targeted disruptions of the IFT172 gene, eliminating functional protein expression and creating a versatile model for studying ciliogenesis and ciliopathy mechanisms. The polyclonal format provides a heterogeneous allele pool suitable for population-based assays without clonal selection bias.
HAP1 is a fibroblast-like human cell line with a near-haploid karyotype, originally from a male chronic myeloid leukemia patient. Its haploidy simplifies gene editing, requiring disruption of a single allele for functional knockout, making it ideal for functional genomics, CRISPR screens, and pathway dissection. HAP1 cells retain ciliary assembly and Hedgehog signaling components, offering a physiologically relevant platform for cilia research.
IFT172 encodes a core IFT-B complex subunit that, together with KIF3A/KIF3B kinesin-2 motors, facilitates anterograde ciliary transport. It physically associates with IFT88, IFT20, IFT57, and IFT-A components, and is critical for delivering Smoothened (SMO) and other cargo. Transcription is driven by RFX factors and FOXJ1, and IFT172 function is required for Hedgehog-dependent activation of GLI transcription factors and expression of PTCH1. Its loss disrupts IFT-B integrity, halting anterograde trafficking and impairing ciliogenesis and downstream signaling, with implications in TGF-?? and cAMP pathways.
Loss-of-function mutations in IFT172 are linked to severe ciliopathies, including Jeune syndrome, Mainzer-Saldino syndrome, nephronophthisis, Bardet-Biedl syndrome, retinitis pigmentosa, and obesity. The HAP1 IFT172 knockout cells faithfully mirror ciliogenesis defects and Hedgehog signaling attenuation observed in patients, establishing them as a valuable disease model. The near-haploid genome facilitates straightforward complementation with wild-type or mutant IFT172 constructs and quantitative assessment of gene dosage effects, supporting mechanistic studies and drug testing.
This knockout population supports immunofluorescence microscopy for cilia markers (acetylated tubulin, ARL13B) to measure cilia length and frequency, and high-content screens for ciliogenesis modulators. Biochemical analyses of IFT-B complex integrity via co-immunoprecipitation and western blotting, along with RT-qPCR of Hedgehog targets (GLI1, PTCH1), are readily performed. The cells enable flow cytometric assessment of ciliary GPCR trafficking and migration/invasion assays, and are adaptable to high-throughput drug discovery for ciliopathies. For further details or custom applications, contact Ascent Research.
