The EIF5A2 Knockout NCI-H1299 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human non-small cell lung carcinoma line NCI-H1299. This product features targeted disruption of the EIF5A2 gene, generating a heterogeneous pool of loss-of-function alleles suitable for studying gene function without clonal selection artifacts. The polyclonal format minimizes single-cell bottleneck effects and clonal adaptation, providing a representative population for downstream assays.
NCI-H1299 is an epithelial cell line isolated from a metastatic lymph node of a lung adenocarcinoma patient. It serves as a well-established model for lung cancer metastasis and oncogenic signaling, recapitulating features of aggressive tumor biology including epithelial-mesenchymal transition (EMT) and invasive migration. The line’s genetic background supports investigations of mTOR, MAPK/ERK, and PI3K/AKT pathway dependencies.
EIF5A2 encodes a translation elongation factor activated by hypusination, a modification catalyzed by deoxyhypusine synthase (DHPS) and deoxyhypusine hydroxylase (DOHH), and regulated by mTORC1. Hypusinated EIF5A2 facilitates selective translation of mRNAs encoding proliferation and metastasis drivers such as cyclin D1, c-MYC, vimentin, and Snail. It interacts with ribosomal proteins RPL5 and RPL11 and is exported by exportin-4 (XPO4). Upstream regulators including MYC and growth factors (EGF, PDGF) position EIF5A2 at the intersection of mTORC1, MAPK/ERK, and PI3K/AKT signaling networks.
In NCI-H1299 cells, EIF5A2 overexpression enhances translation of pro-proliferative and pro-metastatic mRNAs, promoting cell growth, survival, and EMT. The CRISPR/Cas9-mediated knockout in this polyclonal population enables dissection of lung adenocarcinoma dependence on hypusination-driven translation. It allows assessment of how EIF5A2 loss impacts actin cytoskeleton reorganization, adhesion, and the cadherin switch during metastatic progression.
Researchers can employ these knockout cells to examine EIF5A2-dependent translational regulation, probe EMT mechanisms via immunoblotting and RT-qPCR, and perform proliferation (MTT), migration (Boyden chamber), and apoptosis (Annexin V) assays. The model is suitable for co-immunoprecipitation of hypusination factors and screening of DHPS/DOHH inhibitors. Drug sensitivity testing with PI3K/AKT or MEK/ERK inhibitors can identify context-specific vulnerabilities. For further information, please contact Ascent Research.
