The GPR107 Knockout NCI-H1975 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the NCI-H1975 human lung adenocarcinoma cell line. This product comprises a heterogeneous pool of cells carrying CRISPR-mediated disruptions in the GPR107 gene, enabling loss-of-function analysis without clonal isolation. The polyclonal format preserves population diversity and is well-suited for functional assays where pooled responses may reveal phenotypic trends masked in monoclonal lines.
The NCI-H1975 host line is a well-characterized epithelial cell model of non-small cell lung cancer (NSCLC) containing EGFR L858R/T790M mutations. The L858R mutation confers constitutive kinase activity, while T790M reduces affinity for first- and second-generation tyrosine kinase inhibitors, making this line particularly valuable for studying acquired drug resistance and EGFR-driven oncogenic signaling.
GPR107 encodes an orphan G protein-coupled receptor that predominantly localizes to the trans-Golgi network and participates in clathrin-dependent trafficking. Specifically, GPR107 interacts with the AP-1 adaptor complex, clathrin heavy chain, and the GTPase ARF1, positioning it as a key organizer of Golgi-to-endosome transport. Transcriptionally, GPR107 is downstream of p53 and is modulated by EGFR signaling, thereby linking tumor-suppressive and oncogenic networks. Functionally, GPR107 loss alters EGFR degradation kinetics, integrin trafficking, and AP-1?Cdependent sorting of cargoes, impacting receptor signal termination and cell adhesion dynamics.
In the NCI-H1975 background, disruption of GPR107 is predicted to perturb EGFR endocytic recycling and degradation, potentially sustaining downstream kinase activation and attenuating the efficacy of EGFR-targeted therapies. Additionally, altered integrin trafficking may modulate cell migration and invasion, critical determinants of metastatic dissemination in lung adenocarcinoma. Thus, this knockout model provides a platform for interrogating the interplay between Golgi trafficking, receptor turnover, and malignant phenotypes in an EGFR-mutant context.
The polyclonal knockout cells are amenable to a wide range of experimental techniques, including Western blot analysis of EGFR, AKT, and ERK phosphorylation; EGFR degradation assays using cycloheximide or bafilomycin A1; and transferrin uptake assays to measure clathrin-mediated endocytosis. Further applications include cell proliferation and transwell migration/invasion assays, immunofluorescence to visualize Golgi architecture and EGFR subcellular localization, and co-immunoprecipitation to assess GPR107?CAP-1 interactions. Together, these approaches facilitate detailed mechanistic studies of GPCR-mediated trafficking, endocytosis, and drug resistance in NSCLC. For additional information, please contact Ascent Research.
