The APOL2 Knockout HAP1 Polyclonal Cells represent a CRISPR/Cas9-engineered polyclonal knockout population in which the APOL2 gene has been disrupted across a heterogeneous pool of cells. This approach avoids the clonal selection bottlenecks inherent to single?cell derived lines and provides a more robust representation of gene function in a near?haploid background.
The HAP1 parental cell line is a human near-haploid chronic myeloid leukemia (CML)-derived line with an adherent, fibroblast-like morphology. Originally derived from the KBM-7 CML cell line, HAP1 cells retain haploidy for most chromosomes, with the exception of a disomic region on chromosome 15. This near-haploid karyotype simplifies genetic analysis, as most gene disruptions result in a single active allele, making HAP1 an ideal host for CRISPR-based knockout screens and functional genomic studies.
APOL2 encodes a member of the apolipoprotein L family that functions in lipid binding and transport, and critically, in the negative regulation of apoptosis and modulation of autophagy. At the mitochondrial membrane, APOL2 interacts directly with the anti?apoptotic protein BCL2L1, thereby inhibiting caspase activation and promoting cell survival. Additionally, APOL2 binds to BECN1, a key regulator of autophagosome formation, and modulates autophagy flux. Its expression is transcriptionally activated by TNF and IFNG through the NF???B pathway, with NFKB1 acting as a downstream effector. Together, these interactions position APOL2 at the intersection of TNF/NF???B signaling, apoptosis, and autophagy??coordinating cell survival and innate immune responses.
The near-haploid HAP1 background eliminates the confounding effects of a second functional allele, ensuring that CRISPR/Cas9-mediated disruption of APOL2 generates a complete loss-of-function model. This is especially critical for studying APOL2??s dual role in apoptosis inhibition and autophagy regulation, where even partial gene activity can mask phenotypes. The polyclonal knockout format further strengthens the model by minimizing clonal heterogeneity and enabling bulk assays such as drug?response profiling, RNA?sequencing, and proteomic analyses. In the context of CML biology, APOL2 knockout in HAP1 cells provides a defined system to dissect how cytokine?induced anti?apoptotic signaling contributes to leukemia cell survival.
Researchers can employ the APOL2 Knockout HAP1 Polyclonal Cells in a variety of downstream applications, including apoptosis assays by annexin V flow cytometry to quantify TNF?induced cell death, and autophagy flux measurements via LC3 puncta immunostaining or western blotting for BECN1 and BCL2L1 complex formation. Co?immunoprecipitation experiments can validate APOL2 interactions with these effectors, while RNA?seq and RT?qPCR can profile NF???B?mediated transcriptional programs. The cells are also suited for drug target validation studies, particularly for compounds targeting the TNF/NF???B pathway or anti?apoptotic BCL2 family members. For technical specifications or to discuss research applications, please contact Ascent Research.
