The ARL3 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed for the disruption of the ARL3 gene in the HAP1 host cell background. This polyclonal knockout product leverages the HAP1 cell line to provide a versatile model for studying gene function without the need for single-cell cloning, offering a heterogeneous pool of cells with targeted gene disruption. The product is intended for advanced biomedical research applications, enabling the investigation of ARL3-associated biological processes and disease mechanisms in a near-haploid genetic context.
The host cell line, HAP1, is a near-haploid fibroblast-like leukemia cell line derived from the KBM-7 chronic myeloid leukemia line of human male origin. Its haploid karyotype makes it a powerful tool for genetic perturbation studies, as a single allele disruption typically leads to a loss-of-function phenotype, reducing genetic redundancy and simplifying genotype-phenotype correlations. This characteristic renders HAP1 cells particularly suitable for CRISPR-based knockout experiments, functional genomic screens, and signaling pathway dissection in a human cellular context.
ARL3 encodes a small GTPase that acts as a molecular switch in ciliary trafficking, cycling between a GDP-bound inactive state and a GTP-bound active state. Active ARL3 promotes the release of lipidated cargo proteins, such as NPHP3 and INPP5E, from their carrier proteins UNC119 and PDE6D at the ciliary base, facilitating their entry into the cilium. Upstream regulators include the guanine nucleotide exchange factor ARL13B and the GTPase-activating protein RP2, which control ARL3 activation and inactivation, respectively. ARL3 interacts with ARL13B, RP2, UNC119A, PDE6D, and BBSome components, and functions within pathways governing cilium assembly, intracellular protein transport, and photoreceptor cell maintenance. Dysregulation of ARL3-mediated trafficking is implicated in ciliopathies such as retinitis pigmentosa, Joubert syndrome, and Leber congenital amaurosis.
In the HAP1 background, the ARL3 knockout model provides a defined system to dissect ARL3-dependent signaling and ciliary protein networks. The near-haploid nature of HAP1 cells ensures efficient disruption of ARL3 function, eliminating potential confounding effects from a second functional allele. This knockout population enables robust interrogation of ciliary trafficking mechanisms and disease-relevant pathways in a simplified genomic context, making it an ideal platform for studying the molecular pathology of ciliopathies and assessing the role of small GTPase signaling in cancer cell biology.
This polyclonal knockout product supports a wide range of research applications, including the investigation of ciliopathy mechanisms, screening for modulators of ciliary trafficking, functional analysis of photoreceptor protein transport, and studies of small GTPase signaling in leukemia cells. Representative experimental approaches include immunofluorescence staining to assess ciliary structure and protein localization, western blotting to evaluate ARL3 and interacting partner expression, co-immunoprecipitation to detect ARL3 protein complexes, ciliogenesis assays to monitor cilium formation, GTPase activity assays to measure ARL3 activation status, and CRISPR-based rescue experiments to validate phenotype specificity. For additional product specifications or technical support, please contact Ascent Research.
