The AIFM2 Knockout Jurkat Polyclonal Cells product comprises a CRISPR/Cas9-edited polyclonal knockout cell population in which the AIFM2 gene has been disrupted, creating a loss-of-function model for functional genomics. This pooled format retains the genetic diversity characteristic of a polyclonal population, facilitating studies that require representation of multiple editing events without the constraints of monoclonal isolation. The model is specifically designed for investigating ferroptosis regulation and caspase-independent apoptosis in a T-lymphoblast background.
The host Jurkat cell line is an immortalized T lymphoblast derived from the peripheral blood of a 14-year-old male with acute T-cell leukemia. It serves as a well-established model for T-cell receptor signaling, apoptosis, and immunological research. Jurkat cells exhibit rapid proliferation, ease of genetic manipulation, and characterized responses to oxidative stress and death receptor ligands, providing a physiologically relevant context for AIFM2 functional studies.
AIFM2 encodes ferroptosis suppressor protein 1 (FSP1), an NAD(P)H-dependent oxidoreductase that reduces coenzyme Q10 to ubiquinol, thereby trapping lipid peroxyl radicals and inhibiting lipid peroxidation. This mechanism operates in parallel with GPX4 to prevent ferroptosis. AIFM2 expression is regulated by p53 (TP53) and NRF2 (NFE2L2) in response to DNA damage and oxidative stress. Downstream, FSP1 maintains the reduced ubiquinol pool, while under severe apoptotic stimuli, it translocates to the nucleus to mediate caspase-independent DNA fragmentation, interacting with apoptosis-inducing factor (AIF) and endonuclease G.
In Jurkat cells, AIFM2 knockout enables precise interrogation of ferroptosis pathways relevant to T-cell malignancies. The absence of FSP1 potentiates lipid peroxidation-dependent cell death induced by agents such as RSL3 and erastin, making this model valuable for studying tumor cell susceptibility to ferroptosis. It also allows dissection of the interplay between GPX4 and FSP1 systems, and the role of p53/NRF2 signaling in redox homeostasis. The polyclonal setting captures population-level heterogeneity, aiding in understanding clonal responses to ferroptosis induction.
Typical applications include flow cytometric detection of lipid peroxidation with C11-BODIPY, western blotting for GPX4 and ACSL4, glutathione measurements, and caspase activity assays. Co-immunoprecipitation can probe FSP1?CCoQ10 interactions. Ferroptosis induction and rescue experiments with RSL3 or liproxstatin-1, combined with cell viability assays, validate functional dependencies. Mitochondrial membrane potential assessment using JC-1 further links AIFM2 to apoptotic processes. This AIFM2 knockout Jurkat polyclonal cell population thus supports drug screening, mechanistic studies, and immunogenic cell death research. For further information, please contact Ascent Research.
