The APOL1 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HAP1 human cell line, engineered to disrupt the APOL1 gene. This heterogeneous pool of knockout cells provides a robust loss-of-function model for investigating APOL1 biology in innate immunity, lipid metabolism, and kidney disease. The polyclonal format ensures broad allele representation, minimizing clonal bias and enabling more physiologically relevant studies.
HAP1 is a near-haploid human cell line derived from chronic myelogenous leukemia patient KBM-7. Its single copy of most chromosomes simplifies genetic manipulation and functional interpretation, making it widely used for knockout studies and genome-wide screens. This background retains key signaling pathways, supporting research in innate immunity and cell death.
APOL1 encodes a component of high-density lipoprotein (HDL) and functions as a trypanolytic factor in innate immunity. Its expression is activated by interferon-gamma (IFN-??) and tumor necrosis factor-alpha (TNF-??) via JAK-STAT and NF-??B signaling. Downstream, APOL1 mediates trypanosome lysis, while disease-associated G1 and G2 variants induce podocyte dysfunction by promoting mitochondrial membrane permeabilization, cytochrome c release, caspase-3 activation, and lysosomal permeability. APOL1 also interacts with APOA1 and modulates autophagic flux, linking lipid metabolism to programmed cell death.
In the HAP1 near-haploid background, APOL1 knockout enables clear dissection of its roles in innate immunity and cell death without compensatory genetic effects. The line’s leukemic origin provides a straightforward model to study APOL1-dependent mitochondrial and lysosomal dysfunction. Moreover, HAP1 cells express key pathway components such as STAT1, allowing direct investigation of IFN-??-APOL1 signaling axes. This is especially advantageous for high-throughput assays examining APOL1 variant toxicity and therapeutic target screening, as the simplified genome reduces confounding variability.
Applications include evaluating APOL1 trypanolytic activity, investigating variant-induced podocyte injury mechanisms in focal segmental glomerulosclerosis and HIV-associated nephropathy, and screening for modulators of APOL1-mediated cytotoxicity. Typical assays involve western blotting and RT-qPCR for knockout validation, trypanosome lysis assays, caspase-3 activation and Annexin V staining, mitochondrial membrane potential measurements, and autophagic flux analysis. The polyclonal population ensures versatility for reproducible functional studies. For further information, please contact Ascent Research.
