IFT22 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the near-haploid HAP1 cell line, providing a loss-of-function model for the small GTPase IFT22. This polyclonal format enables robust target-gene disruption without clonal isolation, supporting rapid functional screening and pathway analysis in ciliary biology.
The HAP1 host cell line, originating from KBM-7 chronic myeloid leukemia cells, is adherent and fibroblast-like with a near-haploid karyotype ideal for genetic screens. This background facilitates clear interpretation of knockout phenotypes, as single-copy gene disruption unmasks recessive mutations, making it a preferred platform for CRISPR-based functional genomics.
IFT22 is an integral component of the intraflagellar transport (IFT) complex A, essential for retrograde trafficking within primary cilia. It interacts with IFT-A partners IFT43, IFT121, IFT139, and IFT144, as well as IFT70. Upstream regulators include RFX transcription factors and FOXJ1, integrating Hedgehog signaling and cell cycle cues. Downstream, IFT22 mediates retrograde IFT, ciliary disassembly, and Hedgehog pathway output via SMO and GLI1 activation. The knockout disrupts ciliogenesis and interconnected signaling networks involving IFT-B components (IFT88, IFT52), kinesin-2, dynein-2, the BBSome, and transition zone proteins (TCTN1, TMEM216).
In HAP1 cells, IFT22 knockout impairs ciliary assembly and attenuates Hedgehog and Wnt signaling, offering a haploid genetic model to dissect IFT-A function. The polyclonal knockout population is particularly suited for comparing mutant and wild-type phenotypes, facilitating modifier screens and complementation studies. This system helps delineate IFT22??s specific role among IFT-A components, with direct relevance to ciliopathy mechanisms.
Applications include immunofluorescence staining for ciliary markers (acetylated tubulin, ARL13B), western blot analysis of IFT and Hedgehog proteins (GLI1, PTCH1), and RT-qPCR profiling of Hedgehog target genes. Live-cell imaging of IFT dynamics and flow cytometric analysis of ciliated populations are also feasible. This product is suitable for modeling ciliopathies (Joubert syndrome, Meckel syndrome, Bardet-Biedel syndrome), investigating polycystic kidney disease, and screening for therapeutic modulators of ciliary signaling. For further information, contact Ascent Research.
