The KDM6A Knockout NCI-H1975 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population featuring targeted disruption of the KDM6A gene. This product is supplied as a heterogeneous pool of cells, enabling loss-of-function studies without the biases of clonal selection. The polyclonal format preserves genetic diversity, making it particularly suitable for population-level analyses of epigenetic reprogramming and drug sensitivity profiling.
The host NCI-H1975 cell line is a human lung adenocarcinoma model derived from a non-small cell lung cancer patient. These cells harbor an activating EGFR L858R mutation and the secondary T790M gatekeeper mutation, which together confer resistance to the EGFR tyrosine kinase inhibitor gefitinib. The clinically relevant dual-mutation background establishes NCI-H1975 as a widely used system for investigating acquired EGFR-TKI resistance and the biology of lung adenocarcinoma.
KDM6A encodes a histone H3K27 demethylase that removes the repressive H3K27me3 mark, a modification deposited by the Polycomb repressive complex 2 (PRC2) containing EZH2 and SUZ12. By erasing this mark, KDM6A activates transcription of target genes such as HOXA1, CDKN1A (p21), SMAD7, and CDH1 (E-cadherin). KDM6A is recruited to chromatin through interactions with MLL3/MLL4 complexes (including WDR5, ASH2L, RBBP5) and the SWI/SNF remodeling complex, and its expression is directly regulated by retinoic acid receptors, the Notch intracellular domain/RBPJ complex, and TGF-??/SMAD2/3 signaling. Thus, KDM6A integrates multiple developmental and tumor-suppressive inputs to orchestrate gene expression programs.
In the NCI-H1975 model, knockout of KDM6A is expected to increase H3K27me3 occupancy at promoters of tumor suppressors like CDKN1A and CDH1, potentially silencing their expression and enhancing malignant characteristics. Combined with the gefitinib-resistant EGFR L858R/T790M background, loss of KDM6A provides a platform to dissect the epigenetic contribution to drug resistance and to explore synthetic vulnerabilities. This model enables investigation of how histone demethylase inactivation intersects with oncogenic EGFR signaling to drive tumor progression.
This knockout model is well-suited for ChIP-qPCR to map H3K27me3 changes, RNA-seq for transcriptomic profiling, and Western blotting to confirm KDM6A loss and H3K27me3 accumulation. Functional assays such as proliferation, migration/invasion, and gefitinib or demethylase inhibitor dose-response studies can be conducted. Applications include epigenetic regulation studies, Notch pathway analysis, and drug resistance mechanism research. For further information, please contact Ascent Research.
