This product provides a CRISPR/Cas9-edited polyclonal knockout cell population targeting the DIP2A gene in the NCI-H1975 human lung adenocarcinoma cell line. The polyclonal nature of the knockout pool avoids clonal artifacts and enables robust population-level loss-of-function studies without the necessity of single-cell clone isolation. CRISPR/Cas9-mediated gene disruption has been employed to generate a heterogeneous knockout model, which is ideal for functional genomics and drug target evaluation experiments. These cells are supplied as a ready-to-use in vitro model, facilitating immediate experimental application without additional genetic manipulation.
The NCI-H1975 cell line is a human lung adenocarcinoma model derived from a non-smoking female. It harbors EGFR L858R and T790M mutations, making it relevant for tyrosine kinase inhibitor resistance studies. This adherent epithelial line is widely used in NSCLC research to study EGFR-driven signaling and therapeutic responses. Incorporating DIP2A knockout into this genetic background creates a powerful tool for examining the convergence of FSTL1/DIP2A signaling with EGFR pathways.
DIP2A encodes a transmembrane protein that serves as a receptor for FSTL1, a secreted glycoprotein. Ligand binding activates PI3K/AKT/mTOR signaling, driving cell proliferation and survival, and also engages the DCC netrin-1 receptor to modulate axon guidance and migration. DIP2A interacts with SMAD2/3, potentially mediating cross-talk with TGF-??/BMP pathways. Key downstream effectors include AKT1, mTOR, BCL2 family apoptosis regulators, and the Rho GTPases RAC1 and CDC42, which control cytoskeletal dynamics. DIP2A expression is transcriptionally regulated by SP1 and can be silenced by DNA methylation. Thus, DIP2A integrates FSTL1-initiated and DCC-dependent signals, contributing to both developmental processes and oncogenic behaviors.
In NCI-H1975 cells, DIP2A knockout is expected to abrogate FSTL1-induced AKT phosphorylation, impairing PI3K/AKT/mTOR signaling. This loss-of-function model likely reduces cell proliferation, promotes apoptosis via BCL2 family dysregulation, and diminishes migration and invasion through altered RAC1/CDC42 activity. By decoupling DIP2A signaling from EGFR pathway addiction, this model enables dissection of DIP2A??s specific contribution to NSCLC malignancy and exploration of synthetic vulnerabilities.
Key applications include western blotting for DIP2A and phospho-AKT, RT-qPCR for DIP2A mRNA, MTS proliferation assays, Annexin V apoptosis staining, and Transwell migration/invasion assays. Co-immunoprecipitation with DCC or SMAD2/3 can probe protein complexes, while FSTL1 stimulation time courses assess downstream signaling kinetics. These assays support drug target validation, functional genomics, and mechanistic studies of the FSTL1/DIP2A axis in lung adenocarcinoma. For more information, please contact Ascent Research.
