The KDM3B Knockout NCI-H1975 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population generated from the NCI-H1975 human lung adenocarcinoma cell line. This product disrupts the KDM3B gene, creating a heterogeneous pool of cells with targeted gene disruption to study loss-of-function phenotypes in a pooled format. Unlike clonal lines, these polyclonal cells better represent the genetic heterogeneity of tumors and are suitable for bulk functional assays without clonal selection bias. The knockout is achieved through CRISPR/Cas9-mediated gene disruption, enabling researchers to interrogate KDM3B-dependent mechanisms in a non-small cell lung cancer context.
The parental NCI-H1975 cell line is an epithelial cell line derived from a female never-smoker with lung adenocarcinoma. It harbors activating EGFR L858R and T790M mutations, which are clinically relevant drivers of NSCLC and confer sensitivity or resistance to EGFR tyrosine kinase inhibitors (TKIs). NCI-H1975 cells are widely used as a model for investigating EGFR-mutant lung adenocarcinoma, drug resistance, and the tumor microenvironment. The adherent, epithelial morphology of these cells also makes them amenable to standard cell culture and imaging techniques.
KDM3B encodes a histone demethylase that specifically removes mono- and dimethyl marks from lysine 9 of histone H3 (H3K9me1/me2), thereby relieving transcriptional repression and acting as a coactivator. KDM3B is regulated by hypoxia-inducible factor 1-alpha (HIF1A), androgen receptor (AR), and NF-??B, and it interacts directly with AR and NF-??B. By demethylating H3K9me2 at the promoters of target genes, KDM3B facilitates transcriptional activation of pathways controlling cell cycle progression and apoptosis. Representative pathway components include HIF1A, AR, H3K9me2, KDM3B, and downstream target promoters. Loss of KDM3B function therefore disrupts these gene expression programs, altering cellular responses to environmental cues and oncogenic signals.
In the NCI-H1975 lung adenocarcinoma model, KDM3B knockout polyclonal cells provide a powerful tool for dissecting the epigenetic regulation of cancer phenotypes. Because NCI-H1975 cells exhibit EGFR-mutant-driven proliferation and acquired T790M-mediated resistance to first- and second-generation EGFR TKIs, KDM3B loss may modulate transcriptional networks that influence drug sensitivity, cell migration, and survival. The polyclonal nature of this product preserves the complexity of a heterogeneous knockout pool, making it suitable for studying population-level responses to targeted therapies and for identifying genes whose expression is epigenetically controlled by KDM3B in a therapeutically relevant cancer background.
Researchers can employ these knockout cells in a variety of assays, including western blotting for KDM3B and H3K9me2 levels, RT-qPCR and RNA-seq for transcriptomic changes, ChIP-qPCR for H3K9me2 enrichment at specific promoters, and functional assays such as MTT proliferation, Transwell migration, and EGFR TKI drug sensitivity testing. Applications span functional characterization of KDM3B in NSCLC, investigation of epigenetic mechanisms underlying drug resistance, dissection of signaling crosstalk between HIF, AR, and NF-kB pathways, and preclinical screening of epigenetic modifiers. For additional information on product specifications and technical support, please contact Ascent Research.
