The ATP5MJ Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population generated from HEK293T human embryonic kidney cells, targeting the ATP5MJ gene via CRISPR/Cas9-mediated gene disruption. This product provides a heterogeneous pool of cells carrying ATP5MJ loss-of-function mutations, enabling population-level analysis of ATP5MJ-dependent pathways without clonal selection artifacts. It is suitable for applications in mitochondrial biology, functional genomics, and drug discovery.
HEK293T cells, derived from HEK293 by stable integration of the SV40 large T antigen, are widely used for high-efficiency transient transfection and protein expression. The human embryonic kidney epithelial origin preserves active mitochondrial oxidative phosphorylation, making these cells an appropriate host for studying ATP synthase function. Their well-characterized metabolism and easy genetic manipulation support targeted gene disruption studies.
ATP5MJ encodes subunit j of mitochondrial ATP synthase (Complex V), a component of the proton channel in the F0 sector essential for coupling the proton gradient to ATP synthesis. Upstream regulators include PGC-1??, NRF1, ERR??, TFAM, and AMPK; downstream, its function impacts ATP production, mitochondrial membrane potential, and ROS generation. Subunit j interacts with ATP5F1A, ATP5F1B, OSCP, ATP5PO, and other inner membrane proteins to maintain Complex V integrity. Knockout disrupts ATP synthase assembly and proton translocation, impairing oxidative phosphorylation.
In HEK293T cells, ATP5MJ knockout serves as a model for mitochondrial complex V deficiency, recapitulating bioenergetic defects relevant to Leigh syndrome, mitochondrial encephalomyopathy, and neurodegenerative disorders. The retained AMPK?CPGC-1?? signaling axis allows investigation of how ATP synthase dysfunction alters mitochondrial biogenesis and cellular energy status. This model bridges molecular-level Complex V disruption with whole-cell metabolic consequences.
Research applications include Seahorse respirometry for oxygen consumption rates, luciferase-based ATP assays, and JC-1/TMRM membrane potential measurements. The cells are also suitable for Complex V activity assays, western blotting of ATP synthase subunits, immunofluorescence imaging of mitochondrial networks, and RT-qPCR for mitochondrial biogenesis genes. They support metabolic pathway analysis and drug screening for mitochondrial dysfunction. For additional inquiries, please contact Ascent Research.
