DMAC2 Knockout HeLa Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HeLa cervical adenocarcinoma cell line. This product features a targeted disruption of the endogenous DMAC2 gene, enabling loss-of-function studies of this mitochondrial complex I assembly factor. The polyclonal format provides a heterogeneous pool of edited alleles, generated through CRISPR/Cas9-mediated gene disruption, suitable for bulk cell-based assays where clonal homogeneity is not required. These cells serve as a versatile model for investigating the molecular mechanisms underlying respiratory chain complex I biogenesis and function.
The HeLa cell line is an immortalized epithelial cell line originating from a HPV-18 positive cervical adenocarcinoma. HeLa cells exhibit abnormal karyotype and robust, adherent growth conducive to routine culture. Widely used in biomedical research, they offer a well-characterized genetic and metabolic background for studying gene perturbations in cancer biology and organelle function. Their rapid proliferation and stable phenotype support high-throughput screening, while their tumorigenic origin enables investigations into cancer-specific metabolic rewiring.
DMAC2 is a critical subunit of the mitochondrial complex I assembly (MCIA) complex, where it interacts with assembly factors such as NDUFAF1, ECSIT, ACAD9, and TMEM126B to facilitate insertion of the membrane arm of respiratory chain complex I (NADH:ubiquinone oxidoreductase). This process is essential for the stability and activity of complex I, which initiates electron transfer in the oxidative phosphorylation pathway. DMAC2 function is indirectly influenced by upstream regulators of mitochondrial biogenesis, including PGC-1??, NRF1, and TFAM, while its loss impairs downstream targets such as mitochondrial membrane potential, ATP synthase activity, and reactive oxygen species (ROS) production. The knockout disrupts the assembly of supercomplexes containing complex I, ubiquinone, and cytochrome c, ultimately compromising electron transport chain efficiency.
In the HeLa cancer cell context, disruption of DMAC2 recapitulates key features of mitochondrial complex I deficiency, a condition linked to disorders such as Leigh syndrome and mitochondrial encephalopathies. Given the inherent metabolic flexibility of cancer cells, this model provides a platform to explore how loss of complex I assembly reshapes cancer metabolic pathways, including alterations in oxidative phosphorylation, glycolysis, and mitochondrial biogenesis. The DMAC2 knockout cells enable dissection of the molecular switch between oxidative metabolism and alternative energy production that underpins tumor cell survival under metabolic stress, offering insights into potential therapeutic vulnerabilities.
These polyclonal knockout cells are suitable for assays investigating mitochondrial function, including Western blotting for complex I subunits (e.g., NDUFS1, NDUFV1), blue native PAGE for supercomplex assembly, and oxygen consumption rate (OCR) measurement. Additional applications include ATP production assays, ROS detection, cell viability under metabolic stress, NADH oxidation assays, and immunoprecipitation of MCIA components. Researchers studying cancer metabolic plasticity, respiratory chain deficiencies, oxidative stress, or drug targeting of mitochondrial metabolism will find this model valuable. For details, contact Ascent Research.
