The CASP6 Knockout HAP1 Polyclonal Cells product provides a CRISPR/Cas9-mediated polyclonal knockout cell population targeting the human CASP6 gene in the HAP1 haploid cell line. This gene-edited pool is designed for loss-of-function studies, enabling investigation of caspase-6-dependent processes without the constraints of single-cell clonal selection. The polyclonal format retains genetic heterogeneity while uniformly disrupting the target locus, making it suitable for pooled screening and robust functional assays. Researchers can employ this model to dissect apoptosis signaling, neurodegeneration, and cancer-related pathways.
The HAP1 cell line is a near-haploid suspension line derived from the chronic myeloid leukemia (CML) KBM-7 clone, providing a simplified genetic background that facilitates gene-editing and functional genomics. Its haploid nature reduces allelic complexity, allowing efficient generation of knockout models on a clean genetic background. Adapted to suspension culture, HAP1 cells are amenable to high-throughput applications, including flow cytometry-based apoptosis assays, viability screens, and biochemical analyses. This host maintains key apoptotic machinery, making it an appropriate context for studying executioner caspases like caspase-6.
CASP6 encodes caspase-6, an executioner caspase activated downstream of initiator caspases-8 and -9 in both intrinsic and extrinsic apoptotic pathways. Upon activation, caspase-6 cleaves critical substrates including lamin A/C (LMNA), promoting nuclear envelope breakdown and apoptosis. It also processes PARP, KRT18, and disease-related proteins huntingtin and APP, linking its activity to neurodegeneration. Upstream activation by Granzyme B and regulation by XIAP and dimerization further modulate its function. Thus, CASP6 knockout disrupts apoptotic execution and nuclear lamina disassembly.
In the HAP1 background, CASP6 knockout eliminates a central apoptotic executioner, enabling dissection of lamin A/C cleavage and nuclear envelope dynamics under stress. Comparing wild-type and knockout responses to inducers like staurosporine or chemotherapeutics via flow cytometry and caspase activity assays reveals apoptotic kinetics. The haploid background minimizes compensation, enhancing phenotypic clarity, and the CML origin allows investigation of caspase-6 in cancer cell survival and drug resistance.
This polyclonal knockout cell pool supports diverse experimental applications, including mechanistic apoptosis studies, neurodegenerative disease modeling (Alzheimer??s and Huntington??s), cancer biology, and genetic modifier screens. Typical assays include western blotting for caspase-6 and its substrates (e.g., lamin A/C), fluorogenic activity measurements, immunofluorescence for nuclear lamina integrity, RT-qPCR for downstream transcription targets, and cell viability tests following stress induction. The product facilitates drug target validation and functional genomics by providing a reliable loss-of-function platform. For further information, please contact Ascent Research.
