The ARL13B Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population in which the ARL13B gene has been disrupted, providing a loss-of-function model for functional studies of ciliary biology and signaling. This polyclonal pool avoids clonal selection artifacts, offering a genetically heterogeneous system that reflects the complexity of ARL13B-dependent processes across a near-haploid background.
The HAP1 parent cell line is a near-haploid, KBM-7-derived human chronic myeloid leukemia model that retains a single copy of most chromosomes, streamlining genetic manipulation and phenotypic analysis. Its haploid genomic architecture reduces functional redundancy and facilitates unambiguous interpretation of knockout effects, making it an ideal platform for investigating pathways that rely on primary cilium integrity and Hedgehog signaling.
ARL13B encodes a ciliary-localized small GTPase that acts as a molecular switch, cycling between GDP- and GTP-bound states regulated by guanine nucleotide exchange factors and GTPase-activating proteins. It is essential for primary cilium formation and Sonic hedgehog (SHH) signaling, controlling ciliary membrane composition and intraflagellar transport (IFT). ARL13B directly interacts with the IFT-B complex, INPP5E, PDE6D, ARL3, and the BBSome, and is required for proper trafficking of ciliary membrane proteins and subsequent activation of GLI transcription factors downstream of the SHH receptors PTCH1 and SMO. Knockout of ARL13B disrupts ciliogenesis and attenuates SHH-mediated transcriptional responses, linking it to ciliopathy defects such as those seen in Joubert syndrome and retinal dystrophy.
In the HAP1 context, ARL13B knockout impairs primary cilium assembly and Hedgehog signal transduction, providing a physiologically relevant model for the molecular dissection of ciliopathies. The near-haploid genotype ensures that knockout effects are not masked by a second functional allele, enabling direct correlation of genotype to phenotype. This system is valuable for exploring ARL13B function in ciliary trafficking, crosstalk with Wnt signaling, and regulation of GLI-dependent gene expression, with relevance to therapeutic target discovery.
Typical applications include ciliopathy disease modeling, high-content imaging-based ciliogenesis assays, western blotting and RT-qPCR analysis of Hedgehog target genes, Gli-reporter assays for SHH pathway activity, and co-immunoprecipitation studies to probe interactions with IFT-B complex members or BBSome components. The cells are also suited for functional rescue experiments, chemical screening for modulators of ciliary signaling, and validation of candidate therapeutic targets. For further technical details or to discuss customized cell engineering projects, please contact Ascent Research.
