The IGSF8 Knockout NCI-H1975 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population derived from the NCI-H1975 human lung adenocarcinoma epithelial cell line, featuring targeted disruption of the immunoglobulin superfamily member 8 (IGSF8) gene. This product consists of a heterogeneous pool of cells carrying diverse loss-of-function genetic modifications at the IGSF8 locus, generated by ribonucleoprotein-mediated delivery of Cas9 nuclease and a gene-specific guide RNA. The polyclonal knockout format retains the natural cellular heterogeneity of the host line, making it well-suited for population-level functional assays without the artifacts associated with clonal selection.
The parental NCI-H1975 line, derived from a female lung adenocarcinoma patient, harbors EGFR L858R and T790M mutations, conferring constitutive receptor activity and partial kinase inhibitor resistance. As a well-established model of non-small cell lung cancer, it provides a clinically relevant epithelial background for gene knockout studies.
IGSF8, also known as EWI-2, is a cell surface glycoprotein that interacts with tetraspanins CD9, CD81, CD82 and integrin ??3??1, organizing tetraspanin-enriched microdomains (TEMs) on the plasma membrane. Through TEMs, IGSF8 modulates integrin-mediated adhesion, spreading, and migration. In NCI-H1975 cells, IGSF8 acts at the crossroads of integrin and EGFR signaling, with expression regulated by EGF and TGF-??. Downstream, it influences focal adhesion kinase (FAK), Src kinases, and the MAPK/ERK cascade, thereby impacting cytoskeletal reorganization and EGFR trafficking.
Disruption of IGSF8 in the NCI-H1975 background creates a powerful tool to dissect the role of tetraspanin-microdomain organization in EGFR-driven oncogenesis and metastasis. Loss of IGSF8 is anticipated to perturb TEM integrity, altering integrin-mediated adhesion and EGFR membrane compartmentalization, with downstream effects on PI3K/AKT and MAPK signaling. This polyclonal knockout system enables population-level assessment of adhesion-dependent signaling, drug sensitivity, and migratory behavior without clonal selection bias, making it particularly relevant for studying how TEM-associated proteins influence therapeutic responses in EGFR-mutant lung adenocarcinoma.
Researchers can apply this polyclonal IGSF8 knockout pool in a variety of experimental contexts, including Western blotting and immunofluorescence to monitor expression changes in tetraspanins and integrins, co-immunoprecipitation to probe protein interactions, and flow cytometry to quantify surface receptor levels. Functional assays such as cell adhesion, migration, and invasion can be paired with phospho-signaling analysis of EGFR, FAK, and Src to map pathway crosstalk. Moreover, drug sensitivity profiling with EGFR tyrosine kinase inhibitors enables investigation of IGSF8’s role in therapeutic resistance. For further technical details or to place an order, please contact Ascent Research.
