The KDM5D Knockout NCI-H1299 Polyclonal Cells product comprises a CRISPR/Cas9-edited polyclonal knockout cell population engineered to disrupt the KDM5D gene in the human NCI-H1299 non-small cell lung carcinoma cell line. This pooled knockout format provides a heterogeneous collection of loss-of-function alleles, minimizing clonal artifacts and enabling robust, population-level functional analyses. It is an ideal tool for investigating the epigenetic mechanisms driven by KDM5D in cancer biology.
NCI-H1299 cells are derived from a metastatic lung adenocarcinoma of lymph node origin and serve as a well-characterized model for NSCLC metastasis. They exhibit epithelial morphology and are deficient for p53, while harboring a homozygous deletion of the CDKN2A locus, which ablates p16INK4a and p14ARF. Importantly, they retain wild-type KRAS and EGFR, making them particularly relevant for studies of epigenetic drivers in lung adenocarcinoma independent of these common oncogenic mutations.
KDM5D functions as a histone demethylase specific for H3K4me2 and H3K4me3, acting as a transcriptional repressor by removing activating methyl marks from gene promoters. Its activity is influenced by upstream regulators including the SP1 transcription factor, androgen receptor, and promoter CpG methylation, and is integrated through interactions with the REST corepressor complex and associated factors HDAC1, HDAC2, and SIN3A/CoREST. Downstream, KDM5D represses genes such as SNAI2, ZEB1, MMP9, CDKN1A, and BAX, thereby linking its demethylase function to control of epithelial-mesenchymal transition, cell cycle progression, and apoptosis.
Disruption of KDM5D in NCI-H1299 cells results in increased H3K4me3 occupancy at the promoters of migration and apoptosis-related genes, leading to altered transcriptional programs that may impact tumorigenicity. This model enables dissection of KDM5D-mediated epigenetic silencing in a lung adenocarcinoma background defined by CDKN2A loss and p53 deficiency. The polyclonal nature captures a spectrum of mutations, providing a more representative loss-of-function phenotype for mechanistic and pharmacological studies.
The product supports a variety of applications including RNA-seq for transcriptome-wide analysis, ChIP-qPCR for histone modification mapping, Western blotting for protein validation, and functional assays such as Transwell migration, Annexin V apoptosis, colony formation, and drug sensitivity profiling. It is also suited for screening demethylase inhibitors and investigating epigenetic regulation of NSCLC metastasis. For further details or technical support, contact Ascent Research.
