The ARAF Knockout Jurkat Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from Jurkat T lymphoblasts through targeted disruption of the human ARAF gene. This heterogeneous pool provides a loss-of-function model for studying ARAF-dependent signaling without clonal selection, enabling investigation of collective gene disruption effects on MAPK/ERK pathway and T cell biology.
Jurkat cells, originally from an acute T cell leukemia patient, are a well-established model for T lymphocyte signaling and immune response. They possess a functional T cell receptor and robust MAPK/ERK activation upon stimulation, making them ideal for dissecting ARAF’s role in T cell activation, proliferation, and survival.
ARAF is a serine/threonine kinase that transduces signals from activated RAS proteins (HRAS, KRAS, NRAS) and receptor tyrosine kinases (EGFR, FGFR) to the MEK-ERK cascade. It interacts with 14-3-3, HSP90, and KSR scaffolds, phosphorylating MEK1/2, which then activate ERK1/2. Nuclear ERK phosphorylates ELK1, inducing c-FOS and c-JUN to regulate proliferation and apoptosis. ARAF operates within the RAS-RAF-MEK-ERK axis, with crosstalk to PI3K-Akt and Rap1 pathways. CRISPR/Cas9-mediated ARAF disruption uncouples RAS from MEK-ERK signaling, impairing growth factor and stress responses.
In T cells, MAPK/ERK signaling drives TCR-mediated activation, cytokine production, and clonal expansion. ARAF knockout Jurkat cells enable dissection of ARAF-specific functions distinct from BRAF or CRAF, and study of oncogenic MAPK dependencies and drug resistance in leukemia. The model also relates to RASopathies like Noonan syndrome and cardio-facio-cutaneous syndrome.
Applications include MAPK pathway signaling screens, drug resistance assays, and TCR signaling studies. Typical readouts include phospho-ERK western blotting, MEK kinase assays, RT-qPCR for FOS and DUSP, CFSE proliferation flow cytometry, Annexin V apoptosis assays, phospho-kinase arrays, and ERK phosphorylation analysis post-TCR stimulation. For more information, contact Ascent Research.
