The GOLGA7 Knockout NCI-H1975 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population derived from the NCI-H1975 human lung adenocarcinoma epithelial cell line, engineered to disrupt the GOLGA7 gene. This product provides a loss-of-function model for investigating Golgi-dependent cellular processes, particularly those linked to cancer biology and drug resistance. The polyclonal nature of the knockout population offers a broad representation of genetic heterogeneity while maintaining the functional consequences of GOLGA7 ablation, making it suitable for pooled functional screenings or bulk biochemical and imaging-based analyses. The gene disruption is achieved via CRISPR/Cas9-mediated targeting, resulting in abrogation of GOLGA7 protein expression across the population.
The host cell line, NCI-H1975, is a widely used model for non-small cell lung cancer (NSCLC) harboring activating EGFR mutations, specifically L858R in exon 21 and T790M in exon 20. These mutations confer constitutive kinase activity and are associated with acquired resistance to first-generation EGFR tyrosine kinase inhibitors (TKIs). NCI-H1975 cells thus provide a clinically relevant backdrop for studying EGFR signaling dynamics, mechanisms of drug resistance, and the influence of cellular organization on oncogenic pathways. The cells grow as an adherent monolayer and exhibit typical epithelial morphology, enabling straightforward adaptation to high-content imaging and biochemical assays.
GOLGA7 encodes a golgin family protein that localizes to the cytosolic face of the Golgi apparatus, where it contributes to stack integrity and vesicle tethering. GOLGA7 interacts with multiple structural and regulatory factors, including the cis-Golgi matrix protein GOLGA2 (GM130), golgins GOLGA3 and GOLGA4, as well as the small GTPases RAB1A and RAB2A, and components of the COPI coatomer complex. Mechanistically, GOLGA7 functions downstream of ARF GTPases and mitotic kinases such as PLK1, and its activity is influenced by Golgi stress signals. It acts upstream of secretory cargo sorting and modulates the trafficking and localization of transmembrane receptors, notably EGFR. Disruption of GOLGA7 is expected to impair Golgi ribbon organization and intra-Golgi transport, thereby affecting post-translational processing and recycling of EGFR back to the plasma membrane.
In the context of EGFR-mutant NCI-H1975 cells, ablation of GOLGA7 introduces a unique perturbation that may alter the spatial regulation of EGFR signaling. Compromised Golgi integrity could lead to defective glycosylation or missorting of EGFR, potentially reducing surface receptor levels or diverting internalized receptors away from recycling endosomes. This may, in turn, modulate downstream signaling cascades such as AKT and ERK pathways, and could sensitize or further desensitize cells to EGFR inhibitors like osimertinib and gefitinib. The model thus enables dissection of the crosstalk between Golgi structure and oncogenic signaling, offering insights into Golgi-mediated mechanisms that contribute to TKI resistance or synthetic vulnerabilities in NSCLC.
Researchers can employ this polyclonal knockout model to investigate Golgi-dependent regulation of EGFR trafficking using immunofluorescence microscopy for Golgi markers (e.g., GM130) and confocal live-cell tracking of fluorescently tagged EGFR. Functional studies may include western blotting for EGFR, phospho-EGFR, AKT, and ERK activation; cell proliferation assays (MTT, BrdU); and comprehensive drug sensitivity profiling with kinase inhibitors. Flow cytometry for surface EGFR quantification and electron microscopy for Golgi ultrastructure provide complementary readouts. The cells are also suitable for genome-wide CRISPR screens to identify synthetic lethal interactions with GOLGA7 loss or targeted chemical screens for Golgi-toxic compounds. For product inquiries or technical assistance, please contact Ascent Research.
