The ICMT Knockout NCI-H1975 Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal population with targeted disruption of the ICMT gene in the NCI-H1975 human lung adenocarcinoma cell line. As a heterogeneous pool of knockout cells, this model enables robust functional studies of ICMT-dependent biology without clonal selection bias. The live-cell product is ready for expansion and analysis in downstream applications.
NCI-H1975 is a widely used non-small cell lung cancer line derived from lung adenocarcinoma, characterized by EGFR L858R and T790M mutations. These alterations drive constitutive kinase activity and oncogenic signaling, rendering the cells sensitive to EGFR tyrosine kinase inhibitors and providing a clinically relevant platform for investigating drug resistance and targeted therapy mechanisms.
ICMT catalyzes the methyl esterification of isoprenylated CAAX proteins, a critical step in the post-translational processing of GTPases. This methylation facilitates stable membrane anchoring of KRAS, HRAS, NRAS, and RHO-family members including CDC42 and RAC1, enabling their regulated cycling and signal propagation. ICMT acts downstream of prenylation by FTase and GGTase-I and cooperates with the endoprotease RCE1. Knockout of ICMT disrupts proper membrane localization of these GTPases, impairing RAS and RHO signaling cascades that control proliferation, cytoskeletal dynamics, and survival.
In the NCI-H1975 background, which relies on EGFR-driven RAS activation for transformed growth, ICMT loss offers a means to genetically dismantle oncogenic signaling. By ablating GTPase methylation, the knockout cells exhibit reduced RAS-mediated transformation, making them suitable for dissecting crosstalk between EGFR and RAS pathways and for evaluating the therapeutic potential of targeting post-prenylation modifications in lung adenocarcinoma.
This polyclonal ICMT knockout model supports diverse assays, including western blotting for unprenylated proteins, cell proliferation and migration assays, and drug sensitivity profiling. Immunofluorescence can visualize disrupted protein localization, and anchorage-independent growth assays assess tumorigenic capacity. These tools facilitate mechanistic inquiry into protein prenylation and methylation in NSCLC. For inquiries, contact Ascent Research.
