GORASP2 Knockout Jurkat Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the Jurkat human T lymphocyte cell line. This loss-of-function model disrupts the GORASP2 gene, encoding the Golgi reassembly stacking protein 2 (GRASP55), a critical mediator of Golgi organization and secretory pathway function. The polyclonal format provides a heterogeneous pool of edited cells, enabling robust loss-of-function studies without single-cell clonal isolation. These cells exhibit loss of GORASP2 protein and aberrant Golgi morphology, serving as a versatile tool for studying Golgi-dependent processes in a T cell context.
The Jurkat cell line, originally established from the peripheral blood of a 14-year-old boy with acute T cell leukemia, is a widely used immortalized human T lymphocyte model. Jurkat cells recapitulate key aspects of T cell receptor (TCR) signaling, activation, and apoptosis, serving as a foundational system for immunological and cancer research. Their rapid proliferation and well-characterized signaling pathways make them ideal for studying the interplay between intracellular trafficking and immune cell function. The GORASP2 knockout in this background allows dissection of Golgi-mediated trafficking requirements for TCR signaling and leukemic cell biology.
GORASP2 plays a central role in Golgi stacking by tethering adjacent cisternae through interactions with its paralog GORASP1 (GRASP65) and the cis-Golgi matrix protein GOLGA2 (GM130). Its function is regulated by phosphorylation: during mitosis, PLK1 and CDK1 phosphorylate GORASP2, triggering Golgi unstacking, while in interphase, JNK-mediated phosphorylation modulates its activity. Downstream, GORASP2 facilitates trafficking of cargo receptors and secretory components, including the nucleotide sugar transporter SLC35A2 and the small GTPase RAB6A. Through these interactions, GORASP2 ensures efficient protein secretion and links cell cycle machinery to organelle dynamics.
In Jurkat T cells, Golgi organization is essential for polarized secretion of cytokines and surface delivery of receptors. Disruption of GORASP2 impairs Golgi ribbon integrity, leading to defective glycosylation and altered trafficking of transmembrane proteins, including TCR components. This perturbation can compromise TCR signal transduction??affecting calcium flux, ERK activation, and NFAT translocation??and may sensitize cells to apoptosis through Golgi stress pathways. Thus, this knockout model enables systematic investigation of how Golgi defects intersect with T cell activation and leukemogenesis, providing insights into secretory pathway dysregulation in hematopoietic malignancies.
Typical applications include analyzing Golgi-dependent cytokine secretion by ELISA, visualizing Golgi fragmentation via confocal immunofluorescence with antibodies against GM130 or GRASP55, and assessing cell cycle-dependent Golgi disassembly using flow cytometry. The cells are also suitable for co-immunoprecipitation to probe GORASP2 interactions with RAB proteins or microtubule motors, and for high-content screening of Golgi trafficking modulators. Furthermore, they serve as a platform to study crosstalk between apoptosis and Golgi stress in T cell leukemia. For additional technical details, please contact Ascent Research.
