The GORASP2 Knockout NCI-H1975 Polyclonal Cells product offers a CRISPR/Cas9?edited polyclonal knockout cell population derived from the NCI-H1975 human lung adenocarcinoma cell line. This polyclonal pool contains a heterogeneous mixture of cells with disrupted GORASP2 alleles, providing a loss?of?function model of the Golgi reassembly stacking protein 2 (GRASP55) that avoids biases associated with single?clone selection.
The host NCI-H1975 cell line is a well?characterized model of non?small cell lung cancer (NSCLC), originally isolated from a patient with lung adenocarcinoma. This cell line harbors the oncogenic EGFR L858R activating mutation and a loss?of?function TP53 mutation, reflecting key genetic drivers found in a substantial subset of human lung tumors. Such a background renders the cells particularly suitable for studies that connect Golgi biology to EGFR?dependent signaling and tumorigenesis.
GORASP2 encodes GRASP55, a peripheral Golgi matrix protein essential for the stacking of Golgi cisternae and the tethering of transport vesicles. Mechanistically, GRASP55 is regulated by phosphorylation through CDK1 and MAPK1/3, and it acts downstream of EGFR and MAP kinase cascades. It physically associates with GORASP1 (GRASP65), the golgins GOLGA2 and GOLGB1, the small GTPase RAB1A, the ARF1 GTPase, and coatomer complex components such as COPB1. Loss of GORASP2 leads to defects in Golgi structural integrity, impaired integrin trafficking, reduced collagen secretion, and altered unconventional secretion of IL?1??, thereby influencing multiple trafficking routes that intersect with oncogenic signaling.
In the NCI-H1975 lung adenocarcinoma background, GORASP2 knockout is predicted to compromise Golgi organization and secretory competence, affecting processes such as integrin?mediated cell adhesion and migration that are critical for metastatic dissemination. The presence of the EGFR L858R mutation further suggests that disruption of GRASP55 may modulate EGFR signaling output or alter the secretion of tumor?promoting factors, providing a platform to investigate how Golgi?dependent trafficking pathways influence oncogenic signaling, cell motility, and drug sensitivity in lung cancer.
Researchers can employ these polyclonal knockout cells in a variety of experimental setups, including immunofluorescence microscopy using GOLGA2 as a cis?Golgi marker to assess Golgi fragmentation, Western blotting to verify GORASP2 ablation and monitor downstream targets, ELISA?based detection of secreted proteins (e.g., IL?6), wound?healing assays for migration analysis, and co?immunoprecipitation to map GRASP55 interactomes. Key applications range from dissecting Golgi structure?function relationships and unconventional secretion mechanisms to evaluating drug responses and modeling Golgi?dependent adhesion in lung adenocarcinoma. For further details, please contact Ascent Research.
