The DIP2A Knockout NCI-H1299 Polyclonal Cells consist of a CRISPR/Cas9-edited polyclonal population with targeted disruption of the DIP2A gene in the NCI-H1299 human lung carcinoma line. This polyclonal format provides a heterogeneous knockout pool suitable for bulk functional studies without clonal selection, enabling loss-of-function analysis of DIP2A in a non-small cell lung cancer (NSCLC) background.
The NCI-H1299 parental line originates from a lymph node metastasis of a non-small cell lung cancer patient. It displays adherent epithelial morphology and carries a homozygous deletion of TP53 (p53-null), while KRAS and EGFR remain wild-type. This genetic profile renders it valuable for studying p53-independent tumor biology, epithelial-mesenchymal transition, and resistance mechanisms.
DIP2A functions as a receptor for Follistatin-like 1 (FSTL1), activating PI3K/Akt/mTOR signaling via Akt phosphorylation (Ser473) and downstream S6 kinase, and potentiating TGF-beta/SMAD transduction through SMAD2 and SMAD3. This dual pathway engagement promotes expression of the EMT transcription factor Snail, as well as Vimentin, c-Myc, and Cyclin D1. The signaling network incorporates GSK3beta and beta-catenin and is subject to regulation by TGF-beta1, miR-200c, and hypoxia, highlighting DIP2A??s role in cell survival, growth, and mesenchymal transition.
In NCI-H1299 cells, DIP2A knockout allows dissection of its contribution to Akt/mTOR-driven proliferation and EMT independently of p53-mediated apoptosis, given the p53-null background. This is critical for elucidating mechanisms of NSCLC metastasis, as DIP2A-dependent Snail and Vimentin upregulation is a key event. Moreover, the absence of EGFR/KRAS mutations in this line underscores DIP2A??s potential as an alternative oncogenic driver and therapeutic target in specific lung cancer subsets.
Typical applications include Western blotting for p-Akt (Ser473), p-S6K, and EMT markers; RT-qPCR for DIP2A and Snail; Transwell migration and invasion assays; and proliferation analyses (MTT, BrdU). Co-immunoprecipitation can assess DIP2A-FSTL1 binding, while phospho-kinase arrays and RNA-seq enable global signaling and transcriptomic profiling. These cells are ideal for drug screens targeting the FSTL1-DIP2A-PI3K/Akt axis. For further information, contact Ascent Research.
