HCCS Knouckout Jurkat Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population in which the HCCS gene is disrupted within the Jurkat human T lymphocyte line. This product provides a genetically mixed pool of cells carrying targeted gene disruptions, enabling the study of HCCS loss-of-function in a robust and well-characterized immune cell background. The polyclonal format avoids clonal selection artifacts and captures a broad range of editing events, making it suitable for population-level analyses of mitochondrial and apoptotic phenotypes.
Jurkat cells are an extensively utilized immortalized T lymphocyte line originally derived from the peripheral blood of a 14-year-old male with acute T cell leukemia. This cell line is a central model system for investigating T cell receptor signaling, cytokine production, apoptosis, and leukemia biology. Jurkat cells exhibit rapid proliferation, suspension growth, and well-defined signaling cascades, including the extrinsic and intrinsic apoptotic pathways, which are critically dependent on mitochondrial integrity.
HCCS encodes holocytochrome c synthase, an enzyme localized to the mitochondrial intermembrane space that catalyzes the covalent attachment of heme to apocytochrome c, a mandatory step for the maturation of cytochrome c (CYCS). Mature cytochrome c is an essential component of the mitochondrial electron transport chain, shuttling electrons between complex III and complex IV, and also functions as a pro-apoptotic factor when released into the cytoplasm. HCCS expression is transcriptionally regulated by nuclear respiratory factor 1 (NRF1), mitochondrial transcription factor A (TFAM), and the coactivator PPARGC1A (PGC-1??). Disruption of HCCS therefore impairs cytochrome c maturation, leading to deficiencies in mitochondrial respiration and altered responsiveness to apoptotic stimuli through the BCL2 family-regulated intrinsic pathway.
In the Jurkat context, HCCS knockout creates a model that dissects the intersection of mitochondrial metabolism and programmed cell death in T cells. Given the heavy reliance of Jurkat cells on mitochondrial respiration and their sensitivity to apoptosis inducers, loss of HCCS function reveals how impaired heme attachment to cytochrome c perturbs respiratory chain activity and downstream caspase activation. This system is particularly relevant for studying mitochondrial complex III deficiency, X-linked microphthalmia with linear skin defects (MLS) syndrome, and the metabolic adaptations of leukemic cells.
Researchers can employ this knockout pool to perform Seahorse-based mitochondrial respiration assays, quantify cytochrome c holoprotein levels by Western blotting, monitor mitochondrial membrane potential with JC-1 dye, and measure apoptosis kinetics via Annexin V staining. The polyclonal population is ideal for screening functional consequences of HCCS ablation under varying metabolic or stress conditions. It further allows exploration of heme trafficking pathways and the interplay between mitochondrial function and T cell activation or exhaustion. For additional technical details, please contact Ascent Research.
