APPL2 Knockout HAP1 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population derived from the near-haploid human HAP1 cell line. This product consists of a heterogeneous pool of cells carrying targeted disruptions in the APPL2 gene, enabling loss-of-function studies without the isolation of single-cell clones. The polyclonal format preserves population-level diversity while providing efficient gene disruption across the culture.
The HAP1 cell line is a near-haploid human cell line originating from the KBM-7 chronic myeloid leukemia cell line. Its haploid karyotype simplifies gene targeting, as a single successful editing event is sufficient to generate a complete knockout, unlike diploid lines where biallelic targeting is required. This feature makes HAP1 cells a robust platform for functional genomics, particularly in CRISPR-based knockout screens and metabolic or signaling pathway analyses.
APPL2 encodes an adaptor protein that mediates intracellular signaling and endosomal trafficking. It functions downstream of activated receptors such as AdipoR1/R2 and the insulin receptor, and forms complexes with APPL1, Rab5, and PI3K subunits to modulate the PI3K-Akt signaling cascade. Through these interactions, APPL2 regulates key downstream effectors including Akt, GSK3??, and FoxO1, and participates in endosomal receptor sorting. Dysregulation of this network is associated with insulin resistance, metabolic disorders, and cancer, highlighting the importance of APPL2 in cellular homeostasis.
In the HAP1 background, disruption of APPL2 is expected to compromise adapter-mediated signal transduction, particularly metabolic signaling and receptor recycling pathways. The near-haploid state ensures that targeted mutations effectively abolish gene function, creating a clear loss-of-function model. Researchers can use these polyclonal cells to investigate APPL2-dependent endocytic trafficking, insulin and adiponectin signaling, and the resulting effects on cell proliferation and survival, without the need for clonal isolation.
This knockout model is suitable for a variety of functional assays, including Western blotting to assess Akt phosphorylation, immunofluorescence to examine APPL2 subcellular localization, co-immunoprecipitation with Rab5 or PI3K subunits, and flow cytometry for receptor expression. Additional applications encompass glucose uptake assays to probe metabolic effects and migration/invasion assays in cancer research. These APPL2 Knockout HAP1 Polyclonal Cells serve as a versatile tool for drug target validation and mechanistic studies in Type 2 diabetes, obesity, and metabolic syndrome. For further technical information or ordering details, please contact Ascent Research.
