The HTRA2 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from the HAP1 cell line, designed to disrupt the HTRA2 gene. This product offers a heterogeneous pool of cells with targeted gene disruptions, enabling robust loss-of-function studies while avoiding clonal artifacts. Suitable for functional genomics, apoptosis research, and drug discovery, the cells provide a practical tool for investigating HTRA2-dependent mechanisms without the need for clone isolation.
HAP1 is a near-haploid human cell line originating from the KBM-7 chronic myeloid leukemia line. Its haploid karyotype simplifies genetic analysis, as loss-of-function phenotypes are unambiguous without a second allele. Widely used in functional genomics and genetic screening, HAP1 cells retain key signaling pathways relevant to cancer and neurodegeneration, facilitating studies on genes such as HTRA2 that link mitochondrial homeostasis and cell death regulation.
HTRA2 encodes a mitochondrial serine protease that, upon apoptotic stimuli, is released into the cytosol to cleave IAPs like XIAP and cIAP1/2, thereby derepressing caspases and promoting apoptosis. Its activity is regulated by ROS, PINK1 kinase, and PARL protease, which process HTRA2 under mitochondrial stress. HTRA2 also operates within the PINK1-Parkin pathway, interacting directly with PINK1 and Parkin to maintain mitochondrial integrity, and binding Bax and Bak to link mitochondrial permeabilization to caspase activation. Thus, HTRA2 integrates apoptosis signaling with mitochondrial quality control, with implications in cancer and Parkinson??s disease.
In HAP1 cells, HTRA2 knockout eliminates protease activity required for IAP cleavage, impairing apoptosis as measured by reduced caspase-9/3 activation and cytochrome c release. Disruption also affects mitochondrial quality control via PINK1-Parkin, potentially causing accumulation of damaged mitochondria. The haploid background ensures clear phenotypes, and the polyclonal format provides statistically robust data without clonal bias, making it ideal for dissecting apoptosis?Cmitophagy crosstalk.
This polyclonal knockout model is well-suited for advanced research applications. In functional genomics, it enables systematic analysis of HTRA2-dependent networks through co-immunoprecipitation to identify binding partners and Western blotting to assess downstream signaling molecules such as cytochrome c. Apoptosis and mitochondrial dysfunction studies can utilize Annexin V/PI staining, caspase activity assays, and JC-1 mitochondrial membrane potential measurements. The cells also facilitate drug target validation and high-throughput screening of compounds that modulate apoptosis or mitochondrial stress, particularly for cancer and Parkinson??s disease. Genetic modifier screens can be conducted to identify novel regulators of HTRA2 function, leveraging the haploid background for efficient CRISPR-based perturbations. For further information, please contact Ascent Research.
