AMFR Knockout Jurkat Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout pool derived from the Jurkat T lymphoblast line, carrying targeted disruption of the AMFR gene. This mixed population avoids clonal artifacts and enables robust, population-level analysis of AMFR loss-of-function.
Jurkat cells are immortalized human T lymphocytes originally isolated from the peripheral blood of a 14-year-old boy with acute T cell leukemia. They are a standard model for studying T cell receptor signaling, apoptosis, and HIV infection, and their rapid growth and genetic tractability make them ideal for gene editing.
AMFR encodes gp78, an ER-resident RING-finger E3 ubiquitin ligase central to ER-associated degradation (ERAD). gp78 ubiquitinates misfolded proteins??including CD3-delta, apolipoprotein B, and the Wnt co-receptor LRP6??directing them to the proteasome via complexes with VCP/p97, Derlin-1, HRD1, SEL1L, and the E2 enzyme UBE2G2. Under ER stress, gp78 cooperates with UPR sensors IRE1, ATF6, and PERK. Additionally, as the receptor for autocrine motility factor (AMF), gp78 activates migration signaling, modulated by VIMP and UBXD8.
In Jurkat T lymphoblasts, AMFR knockout allows dissection of gp78??s role in TCR subunit degradation; loss of gp78 may stabilize CD3-delta and alter signal transduction. The model is valuable for studying how ERAD impacts HIV replication and T cell survival under apoptotic stimuli, and for isolating AMF-mediated motility signaling relevant to leukemic cell migration.
Applications include ER stress induction with tunicamycin or thapsigargin, Western blotting, RT-qPCR, Transwell migration assays, and flow cytometry for surface marker analysis. Co-immunoprecipitation and in vitro ubiquitination assays characterize gp78 interactions. The knockout cells support research on drug resistance, neurodegeneration models involving ataxin-3 and CFTR deltaF508, and proteasome inhibitor sensitivity. For further details, contact Ascent Research.
