The DMAC2 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the human DMAC2 gene in HEK293T cells. This mixed population provides a versatile loss-of-function model for investigating mitochondrial complex I assembly and related processes. By introducing target-gene disruption across the cell pool, the product enables robust downstream functional studies without the need for single-cell cloning. The polyclonal format preserves biological heterogeneity while ensuring effective reduction of DMAC2-dependent functions, making it suitable for a wide range of experimental applications in molecular and cellular biology.
HEK293T cells are a well-established human embryonic kidney epithelial cell line derived from HEK293 cells and transformed with adenovirus 5 DNA. They constitutively express the SV40 large T antigen, which permits episomal replication of plasmids containing the SV40 origin of replication, resulting in high protein expression and efficient virus production. Their rapid growth, ease of transfection, and robust metabolic activity make them an ideal host for studying mitochondrial biology. The HEK293T background provides a reliable platform for examining the consequences of DMAC2 knockout on oxidative phosphorylation and mitochondrial homeostasis.
DMAC2 (Distal Membrane Arm Assembly Component 2) is a critical mitochondrial complex I assembly factor that stabilizes the ND1 module during biogenesis of the NADH:ubiquinone oxidoreductase complex. It interacts directly with DMAC1, NDUFAF1, NDUFAF2, ND1, and TIMMDC1 to facilitate proper incorporation of the ND1 subunit into complex I. Upstream regulators such as NRF1, TFAM, and PGC-1?? transcriptionally control DMAC2 expression, linking mitochondrial biogenesis to cellular energy demands. DMAC2 knockout leads to destabilization of the ND1 module, impairing NADH:ubiquinone oxidoreductase activity, reducing electron transport, decreasing ATP synthesis, and potentially increasing reactive oxygen species (ROS) production. This disruption affects downstream targets including mitochondrial membrane potential and oxidative phosphorylation efficiency.
In the HEK293T cellular context, loss of DMAC2 function provides a powerful model to dissect mitochondrial complex I assembly pathways. HEK293T cells express the full complement of mitochondrial respiratory chain components, enabling clear phenotypic readouts upon DMAC2 disruption. The knockout population allows researchers to explore the molecular consequences of impaired complex I activity without complications from primary cell variability. This system is particularly valuable for modeling mitochondrial complex I deficiency disorders, such as Leigh syndrome and mitochondrial encephalomyopathy, and for studying metabolic reprogramming in cancer or neurodegeneration where mitochondrial function is altered.
This knockout product supports a wide array of research applications, including mechanistic studies of complex I assembly, screening of therapeutic compounds that target mitochondrial dysfunction, and investigation of metabolic adaptations. Representative assays compatible with these cells include western blotting for complex I subunits, blue-native PAGE to assess mitochondrial supercomplexes, Seahorse respirometry for oxygen consumption rate (OCR) measurement, NADH dehydrogenase activity assays, ROS detection with DCFH-DA, ATP luciferase assays, RT-qPCR for mitochondrial-encoded genes, and immunofluorescence using MitoTracker for mitochondrial morphology. For further information or to acquire this product, please contact Ascent Research.
