The ATPAF2 Knockout Jurkat Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the Jurkat human T-lymphocyte line. This product features targeted gene disruption of the ATPAF2 locus, generating a loss-of-function model that eliminates expression of the mitochondrial F1-ATPase assembly factor.
These polyclonal cells provide a heterogeneous knockout population suitable for investigating mitochondrial Complex V biology and related disorders without clonal selection artifacts.
Jurkat cells are an immortalized suspension lymphoblastoid cell line originally established from the peripheral blood of an acute T-cell leukemia patient. As a widely used T-lymphocyte model, they are extensively employed to study T cell receptor signaling, apoptotic pathways, and HIV infection.
Their well-characterized metabolic profile and ease of genetic manipulation make them a robust host for examining mitochondrial gene function in an immune cell context, particularly the interplay between bioenergetics and T cell effector responses.
The ATPAF2 gene product functions as an essential assembly factor for the F1 catalytic domain of mitochondrial ATP synthase (Complex V). ATPAF2 is regulated by mitochondrial biogenesis factors such as PGC-1?? and NRF1, and participates in the coordinated assembly of Complex V by interacting with ATPAF1 and core F1 subunits, including ATP5A1 and ATP5B.
Loss of ATPAF2 disrupts this assembly process, leading to impaired incorporation of F1 components, defective ATP synthase activity, and subsequent reductions in oxidative phosphorylation, ATP production, and mitochondrial membrane potential.
In Jurkat T cells, mitochondrial ATP generation is critical for sustaining activation, proliferation, and survival signaling. Therefore, ATPAF2 knockout in this cellular environment directly compromises energy metabolism, providing a relevant model to dissect the consequences of Complex V deficiency on T cell function.
It enables the study of how mitochondrial dysfunction intersects with TCR signaling, apoptosis regulation, and metabolic reprogramming, which are central to both normal immunity and leukemogenesis.
Research applications encompass detailed biochemical and functional analyses. Researchers can assess Complex V assembly status via blue native PAGE and immunoblotting for ATP5A1/ATP5B, measure oxygen consumption rate using Seahorse metabolic flux analysis, quantify cellular ATP levels, and evaluate mitochondrial membrane potential with JC-1 staining. The model also supports flow cytometric assessments of mitochondrial mass (MitoTracker) and viability assays under metabolic stress. These cells are valuable for drug screening targeting mitochondrial disorders and for investigating mitochondrial roles in T cell pathologies. For further information, please contact Ascent Research.
