The ATP5PO Knockout HEK293T Polyclonal Cells product provides a heterogeneous cell population derived from HEK293T cells after CRISPR/Cas9-mediated disruption of the ATP5PO gene. This polyclonal knockout pool consists of a mixture of edited alleles, offering a loss-of-function model for studying the OSCP subunit of mitochondrial ATP synthase without clonal selection. The CRISPR/Cas9 approach enables targeted gene disruption, making these cells suitable for examining ATP5PO-dependent mitochondrial functions.
HEK293T cells, a human embryonic kidney cell line, are widely utilized in biomedical research due to their high transfectability and robust protein expression capabilities. Derived from HEK293 cells, they stably express the SV40 large T antigen, which facilitates episomal replication of transfected plasmids. These characteristics make HEK293T an ideal host for generating knockout models, allowing efficient delivery of CRISPR components and straightforward analysis of resulting mitochondrial phenotypes.
ATP5PO encodes the oligomycin sensitivity conferral protein (OSCP), a peripheral stalk subunit of mitochondrial ATP synthase (Complex V). OSCP is critical for coupling the proton motive force generated by the electron transport chain to ATP synthesis. It interacts with core subunits ATP5F1, ATP5F1B, and ATP5F1C, as well as the inhibitory factor ATP5IF1, to stabilize the F1 catalytic domain. The gene is transcriptionally regulated by PPARGC1A, NRF1, and TFAM, key coordinators of mitochondrial biogenesis, and is responsive to thyroid hormone signaling. Disrupting ATP5PO impairs ATP synthase assembly, reduces ATP production, and diminishes mitochondrial membrane potential, leading to compensatory AMPK activation.
Within HEK293T cells, ATP5PO knockout recreates mitochondrial Complex V deficiency, a hallmark of disorders such as Leigh syndrome and mitochondrial encephalomyopathy with lactic acidosis. The polyclonal population recapitulates the energetic stress associated with oxidative phosphorylation failure, making it a relevant model for studying disease mechanisms and metabolic reprogramming. Because HEK293T cells rely on both glycolytic and oxidative metabolism, the knockout allows researchers to dissect the shift toward glycolysis and assess mitochondrial dysfunction under controlled experimental conditions.
These polyclonal knockout cells are suited for a range of experimental applications, including mitochondrial bioenergetics studies using Seahorse respirometry to measure oxygen consumption rate (OCR), ATP luminescence assays to quantify cellular energy status, and JC-1 assays to evaluate mitochondrial membrane potential. They can be employed in drug screening for ATP synthase modulators, metabolic flux analysis, and apoptosis assays such as Annexin V staining to investigate cell fate under energy stress. Additionally, Western blotting can confirm loss of ATP5PO protein and downstream AMPK signaling alterations. For further details or custom inquiry, please contact Ascent Research.
