ISOC1 Knockout HAP1 Polyclonal Cells are a Homo sapiens CRISPR/Cas9-edited polyclonal knockout population derived from the HAP1 near-haploid cell line, designed to disrupt the ISOC1 gene. This loss-of-function model provides researchers with a powerful tool to investigate the biological role of ISOC1, a mitochondrial protein featuring a predicted isochorismatase domain associated with hydrolytic activity. The polyclonal format encompasses a heterogeneous mix of independent editing events, minimizing clonal bias and amenable to pooled screening strategies.
HAP1 is a near-haploid human male cell line originating from the KBM-7 chronic myeloid leukemia lineage, characterized by a predominantly haploid karyotype. This genetic simplicity enables efficient CRISPR/Cas9-mediated gene knockout by targeting a single allele, often resulting in complete functional disruption. HAP1 cells are widely adopted in functional genomics, large-scale knockout screens, and haploid genetic screens because they expose recessive phenotypes without diploid compensation. They also retain key features of leukemic cells, making them a relevant model for oncology and metabolic research.
ISOC1 encodes a protein that localizes to mitochondria and contains an isochorismatase-like domain, a fold commonly found in hydrolases that act on small molecules in cofactor biosynthesis or metabolite salvage. Despite these structural predictions, the specific enzymatic activity, substrate specificity, and biological function of ISOC1 remain uncharacterized. No upstream regulatory signals, downstream effector pathways, or physical interaction partners have been identified. Consequently, the molecular network and signaling context of ISOC1 are undefined, and its knockout is expected to help uncover its cellular roles through systematic functional profiling.
The combination of ISOC1 knockout with the HAP1 haploid background yields a robust experimental system for functional interrogation. The near-haploid genome ensures that CRISPR/Cas9-mediated gene disruption efficiently abrogates ISOC1 function, reducing concerns over residual wild-type alleles. This model is especially suited for metabolic flux analyses under perturbed conditions, such as Seahorse-based respirometry, and for viability screens in the presence of metabolic inhibitors or chemotherapeutics. The leukemic origin of HAP1 cells further permits the investigation of mitochondrial vulnerabilities in the context of cancer metabolism, potentially revealing targets for synthetic lethality.
This polyclonal knockout product supports a wide range of applications, including pooled CRISPR genetic screens to identify synthetic lethality partners, transcriptomic profiling by RNA-seq to map downstream gene expression changes, and mitochondrial functional assays. Knockout validation can be performed using RT-qPCR and western blotting, while immunofluorescence confirms mitochondrial localization of the wild-type protein. High-throughput viability assays can assess sensitivity to mitochondrial stressors or anti-leukemic agents. Overall, ISOC1 Knockout HAP1 Polyclonal Cells provide a versatile and genetically clean platform for dissecting ISOC1 function in mitochondrial biology and disease. For further inquiries, please contact Ascent Research.
