The HERPUD1 Knockout Jurkat Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from the Jurkat human T lymphocyte line. This model disrupts the HERPUD1 gene via Cas9 nuclease activity, creating a heterogeneous loss-of-function system to study endoplasmic reticulum (ER) stress signaling, ER-associated degradation (ERAD), and calcium homeostasis. The polyclonal composition ensures diverse mutational outcomes, providing a robust platform for population-level analyses without clonal bias.
Jurkat cells are an immortalized T lymphocyte line established from a 14-year-old male with acute T cell leukemia. They express a functional T cell receptor (TCR) and secrete interleukin-2 (IL-2) upon activation, serving as a classic model for T cell activation, signaling, and apoptosis. Their well-characterized signal transduction pathways and suspension growth make them ideal for mechanistic studies in cancer and immunology, particularly in the context of TCR-dependent responses and leukemogenesis.
HERPUD1 is an ER-resident protein upregulated by ER stress via ATF6 and spliced XBP1. It facilitates ERAD by bridging ubiquitinated substrates to the 26S proteasome through interactions with VCP/p97, HRD1, and PSMC6, thereby attenuating UPR signaling mediated by IRE1, PERK, and ATF6. HERPUD1 also modulates calcium signaling by binding IP3 receptors (ITPR1), influencing Ca2? flux and affecting downstream targets such as CD3?? and TCR?? efficiency. Upstream stressors include thapsigargin and tunicamycin.
In Jurkat T cells, HERPUD1 disruption impacts ER stress response and TCR-mediated activation. TCR engagement triggers calcium mobilization and UPR, and HERPUD1 loss may alter activation thresholds, IL-2 production, and apoptosis via disrupted ERAD and calcium handling. Interaction with CD3?? implies potential roles in TCR complex assembly. This model enables dissection of survival mechanisms in leukemic T cells, where UPR pathways are often hyperactivated.
This polyclonal knockout cell population supports diverse assays including Western blotting of UPR markers (BiP, CHOP, ATF4), RT-qPCR for ER stress gene profiling, flow cytometry for apoptosis (Annexin V), and calcium flux measurements using Fura-2. Co-immunoprecipitation of HERPUD1 with VCP/p97 or ITPR1 and proteasome activity assays further enable ERAD and calcium signaling studies. Applications extend to drug sensitivity screens targeting proteostasis. For further information, please contact Ascent Research.
