The ARL15 Knockout HAP1 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population targeting the human ARL15 gene. Generated using the near-haploid HAP1 cell line, this product provides a heterogeneous pool of cells with diverse disruption events, ideal for functional genomics studies without clonal bias. The polyclonal format ensures robust representation of loss-of-function alleles, facilitating reliable phenotypic assessments in insulin signaling and metabolic research. These cryopreserved cells are ready for expansion and downstream assay implementation.
HAP1 cells are a human fibroblast-like, near-haploid cell line originally derived from the KBM-7 chronic myeloid leukemia cell line. Their haploidy simplifies CRISPR-mediated gene disruption, as only one allele requires modification to achieve functional knockout, minimizing genetic redundancy and off-target complexities. Extensively used in genetic screening platforms, HAP1 cells offer stable growth kinetics, high transfection efficiency, and compatibility with high-content imaging and automated liquid handling systems. The ARL15 knockout polyclonal population has been validated by genomic DNA PCR and Sanger sequencing to confirm target-gene disruption.
ARL15 encodes a GTPase integral to insulin-stimulated intracellular trafficking and signaling. Upon insulin receptor activation, ARL15 recruits ARNO (CYTH2) to the receptor, promoting endocytosis and enhancing downstream signaling through IRS1, PI3K, and Akt. Active Akt phosphorylates AS160, leading to GLUT4 translocation and glucose uptake. ARL15 thus couples receptor internalization to insulin metabolic responses, with direct relevance to type 2 diabetes and metabolic syndrome.
In the HAP1 model, ARL15 knockout provides a clean genetic background to dissect its specific contributions to insulin pathway dynamics. The near-haploid state eliminates heterozygosity confounders, enabling precise quantification of phospho-signaling events. While native GLUT4 expression is limited, ectopic expression or reporter systems allow detailed analysis of trafficking. This system is particularly suited for studying endocytic routing and Akt-dependent signaling in a simplified cellular context. These features make the model advantageous for mechanistic studies of insulin resistance and type 2 diabetes pathology.
Key applications include functional genomic screens, mechanistic studies of type 2 diabetes, and insulin signaling pathway dissection. Assays such as western blotting for ARL15 and phospho-Akt, RT-qPCR, immunofluorescence for GLUT4 localization, and 2-deoxyglucose uptake assays are commonly employed. Co-immunoprecipitation validates insulin receptor?CARL15 interactions. Genomic DNA PCR and Sanger sequencing confirm knockout. These cells enable high-throughput screening of signaling modulators and detailed characterization of ARL15-dependent phenotypes in metabolic research. For further information, please contact Ascent Research.
