The E2F4 Knockout NCI-H1299 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population designed for loss-of-function studies of the E2F4 transcription factor. This product is generated through CRISPR/Cas9-mediated gene disruption of the E2F4 gene in the NCI-H1299 cell line, creating a heterogeneous pool of knockout cells. As a polyclonal population, it provides a robust model for investigating E2F4-dependent phenotypes without the selective pressure of single-cell cloning, thereby maintaining a broader representation of genetic variability resulting from the editing process.
The NCI-H1299 host cell line is a well-established human non-small cell lung cancer (NSCLC) model derived from the lymph node metastasis of a lung adenocarcinoma from a 43-year-old male. This cell line is widely utilized in cancer biology research due to its reproducible growth characteristics and its relevance to studying the molecular mechanisms underlying lung adenocarcinoma. Its p53-deficient background and other genomic alterations make it a useful system for evaluating tumor suppressor functions and oncogenic signaling pathways.
E2F4 is a transcriptional repressor in the pRB-E2F pathway that, together with DP1/DP2, binds E2F-responsive promoters and recruits HDAC1/SIN3A corepressors to silence S-phase genes. Its repressive activity is controlled by pocket proteins pRB, p107, and p130, themselves phosphorylated by Cyclin D1/CDK4, and by upstream TGF-beta and PI3K/AKT signals. Key downstream targets whose expression is repressed include CCNA2, CCNE1, MYC, CDC6, DHFR, and TK1.
In the context of NCI-H1299 lung cancer cells, E2F4 knockout is predicted to relieve transcriptional repression of proliferation-associated genes, potentially leading to uncontrolled S-phase entry and enhanced cell growth. This model is particularly valuable for dissecting the tumor-suppressive roles of E2F4 in NSCLC, where its loss or functional inactivation has been implicated in disease progression. The polyclonal knockout population allows for the assessment of E2F4-dependent alterations in cell cycle distribution, apoptosis sensitivity, and response to chemotherapeutic agents, providing insights into the role of the pRB-E2F axis in lung adenocarcinoma maintenance.
Researchers can use this knockout model for cell cycle studies, tumor suppressor analysis, and drug target validation. Compatible assays include western blotting for E2F4 and targets, RT-qPCR for cell cycle genes, proliferation assays (MTT, EdU), flow cytometry for cell cycle distribution, colony formation assays, and RNA-seq profiling. The polyclonal population is suitable for functional genomics screens and for testing therapeutic strategies in lung cancer. For further information, please contact Ascent Research.
