The KDM6B Knockout A-549 Polyclonal Cells product offers a CRISPR/Cas9-mediated polyclonal knockout cell population with disruption of the KDM6B gene in the A-549 human lung adenocarcinoma epithelial line. This ready-to-use loss-of-function model enables systematic investigation of KDM6B-dependent epigenetic and transcriptional processes in a population context, avoiding clonal selection bias.
The A-549 host cell line is an adherent epithelial culture established from a lung carcinoma of a 58-year-old Caucasian male. It is a widely accepted model for lung adenocarcinoma and alveolar type II epithelium, exhibiting key features of transformed lung cells and responsiveness to diverse signaling cues, making it suitable for cancer biology and drug discovery studies.
KDM6B (JMJD3) functions as a histone H3K27me3/me2 demethylase, catalyzing the removal of repressive methyl marks and thereby promoting chromatin opening and gene activation. Its expression is induced by upstream regulators such as STAT3, NF-??B, TGF-??/SMAD2/3, retinoic acid receptors, and Notch1 intracellular domain, tying it to TGF-??, NF-??B, and Notch pathways. Downstream, KDM6B transcriptionally activates targets including CDKN2A, CDKN1A, HOX genes, CDH1, and BMP2, and it interacts with partners like the MLL2/3 complex, ASH2L, p53, and BRG1. Knockout of KDM6B in A-549 cells disrupts H3K27me3 demethylation, leading to altered expression of genes governing proliferation, differentiation, senescence, and inflammatory responses.
In the A-549 lung adenocarcinoma model, KDM6B loss provides a tool to dissect how epigenetic deregulation contributes to tumor suppressor silencing, aberrant cell growth, and inflammatory signaling. The interaction with pathways frequently mutated in lung cancer, such as p53 and TGF-??/SMAD, underscores its utility for studying oncogenic transformations and therapeutic vulnerabilities.
Routine applications include western blotting for global H3K27me3 levels, RT-qPCR and ChIP-qPCR to assess expression and histone marks at target loci (e.g., CDKN2A, HOXA), immunofluorescence for subnuclear H3K27me3 distribution, and flow cytometry to quantify apoptosis and cell cycle alterations. Transcriptome-wide RNA-seq can identify novel KDM6B-regulated networks. Functional proliferation and viability measurements use MTT and colony formation assays. This polyclonal knockout cell population supports epigenetics, lung cancer, differentiation, and drug resistance research. For technical inquiries, contact Ascent Research.
