The IFT43 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the near-haploid human HAP1 cell line. These cells provide a loss-of-function model for IFT43, a gene encoding an essential subunit of the intraflagellar transport complex A (IFT-A). The polyclonal population contains a heterogeneous mixture of gene-disrupted cells, enabling robust phenotypic assessment without clonal selection biases, and is particularly useful for pooled screening applications.
HAP1 is a human near-haploid cell line derived from the chronic myeloid leukemia cell line KBM-7. Its haploid karyotype simplifies genetic manipulation, as a single CRISPR/Cas9 targeting event is sufficient to disrupt both alleles in the majority of loci, eliminating confounding effects from wild-type alleles. HAP1 cells retain many epithelial features and are widely used for genetic screens, drug target validation, and signaling pathway dissection.
IFT43 functions as a core component of the IFT-A complex, responsible for retrograde ciliary transport. It interacts with IFT-A subunits including IFT121/WDR35, IFT122, IFT139, IFT140, and IFT144, and is regulated by Aurora A kinase and HDAC6. IFT43 activity is critical for Hedgehog signaling, operating downstream of PTCH1 and SMO to allow proper activation of GLI transcription factors. Additionally, IFT43 contributes to trafficking of the BBSome and ciliary membrane proteins, linking it to ciliogenesis and other signaling networks.
Disruption of IFT43 in the HAP1 background creates a powerful model for ciliopathy research. Loss of IFT43 is associated with Sensenbrenner syndrome, short-rib thoracic dysplasia, retinitis pigmentosa, and nephronophthisis. The near-haploid HAP1 environment enables clean phenotypic readouts of ciliary defects, Hedgehog pathway attenuation, and impaired retrograde transport, facilitating genotype-phenotype correlations that would be masked in diploid systems.
These polyclonal knockout cells are suitable for immunofluorescence staining of cilia, western blotting of IFT components, and RT-qPCR analysis of GLI target expression. Hedgehog reporter assays and proximity ligation assays can map signaling activity and protein interactions. Flow cytometry quantifies ciliated cells, and drug sensitivity screening can identify compounds modulating ciliary trafficking. The polyclonal format also supports functional genomics screens for novel regulators of IFT-A function. For additional information or technical support, please contact Ascent Research.
