The DIABLO Knockout HAP1 Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal population generated from the HAP1 cell line, in which the DIABLO gene (encoding SMAC) has been targeted for disruption. This product provides a heterogeneous knockout pool, enabling unbiased loss-of-function analysis of DIABLO in apoptosis regulation. The polyclonal format is particularly useful for pooled genetic screening applications, as it avoids the limitations of clonal variation and maintains representation of diverse editing events.
HAP1 is a chronic myeloid leukemia-derived cell line with a near-haploid karyotype and stable expression of the BCR-ABL1 fusion kinase. Its near-haploid genome simplifies functional genomics studies by reducing gene redundancy, facilitating robust identification of genotype-phenotype relationships. The oncogenic BCR-ABL1 signaling axis creates a pro-survival cellular milieu, making it a relevant system for investigating mechanisms of apoptosis evasion and therapeutic resistance.
DIABLO (also known as SMAC) is a resident mitochondrial intermembrane protein that functions as a pivotal antagonist of inhibitor of apoptosis proteins (IAPs). Upon receipt of apoptotic stimuli, DIABLO is released into the cytosol, where it directly interacts with XIAP, cIAP1, and cIAP2, displacing them from caspase-9, -3, and -7 and relieving their inhibitory effects. Upstream, its release is orchestrated by pro-apoptotic BAX and BAK, downstream of p53-mediated transcriptional programs, which trigger mitochondrial outer membrane permeabilization and cytochrome c release. Thus, DIABLO serves as a critical molecular link coupling mitochondrial damage to the execution phase of apoptosis.
In the HAP1 background, knockout of DIABLO creates a powerful tool to interrogate IAP-dependent survival signaling that cooperates with BCR-ABL1. This model is particularly valuable for studying how cancer cells subvert mitochondrial apoptosis and for evaluating the efficacy of IAP antagonists (such as Smac mimetics) in conjunction with tyrosine kinase inhibitors. The near-haploid nature further enables high-confidence synthetic lethality screens to discover genes that become essential in the absence of DIABLO, informing combination therapy strategies for myeloid malignancies.
This polyclonal knockout cell product supports a wide range of biochemical and cell-based assays, including western blotting and RT-qPCR for target confirmation, annexin V and caspase-3 cleavage assays for apoptosis quantification, and co-immunoprecipitation to assess DIABLO-XIAP complexes. Additional applications include cytochrome c release detection, mitochondrial membrane potential analysis by flow cytometry, and chemosensitivity testing with agents that trigger intrinsic apoptosis. Researchers can also apply these cells in arrayed or pooled CRISPR modifier screens to identify novel apoptosis regulators. For further technical details and ordering information, please contact Ascent Research.
