The EIF4A2 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 offers a loss-of-function model for studying the EIF4A2 RNA helicase in translational control and cancer biology. The polyclonal pool comprises a heterogeneous mix of edited cells, avoiding clonal selection artifacts and reflecting bulk gene disruption.
The parental NCI-H1299 cell line, established from a lymph node metastasis of lung adenocarcinoma, is an epithelial model of non-small cell lung cancer (NSCLC) with p53 deficiency and wild-type KRAS. It is widely employed to investigate NSCLC progression, metastatic potential, and drug responses. Its metastatic origin and epithelial morphology make it ideal for assays of migration, invasion, and anchorage-independent growth.
EIF4A2 encodes an ATP-dependent DEAD-box RNA helicase integral to the eIF4F translation initiation complex, where it interacts with eIF4G, eIF4E, and cofactors eIF4B and eIF4H. It unwinds 5?? UTR secondary structures to enable ribosomal scanning and cap-dependent translation. Its activity is controlled by mTORC1 signaling, which integrates inputs from growth factor receptors like EGFR, the PI3K/AKT pathway, amino acids, and energy status (AMPK). mTORC1 promotes eIF4E release from 4E-BP and modulates S6K and PDCD4, thereby enhancing translation of structured 5?? UTR mRNAs such as cyclin D1, c-MYC, BCL-2, and VEGF.
In NCI-H1299 cells, EIF4A2 knockout helps dissect how eIF4F-dependent translation drives proliferation, survival, and metastasis. Given the p53-null and KRAS wild-type background, translational control may be critical for oncogenesis. Disrupting EIF4A2 reduces translation of pro-oncogenic factors, offering insights into mTORC1?CeIF4F signaling in NSCLC and evaluating EIF4A2 as a therapeutic target in p53-deficient lung adenocarcinoma.
This polyclonal knockout population supports polysome profiling, ribosome footprinting, and luciferase reporter assays with structured 5?? UTRs to measure cap-dependent translation. It is suitable for western blotting and RT-qPCR of targets like cyclin D1 and c-MYC, co-immunoprecipitation of eIF4F components, and functional assays such as proliferation, migration, and invasion. Drug sensitivity studies with translation inhibitors (e.g., silvestrol, rocaglates) enable target validation. For additional product information, contact Ascent Research.
