The DNMT3A Knockout KYSE-150 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the KYSE-150 human esophageal squamous cell carcinoma line, featuring targeted disruption of the DNMT3A gene. This heterogeneous pool of loss-of-function cells circumvents the limitations of single-cell cloning, preserving genetic diversity that better reflects tumor heterogeneity. By ablating DNMT3A, this product provides a robust tool for investigating de novo DNA methylation-dependent gene silencing in a cancer-relevant context.
KYSE-150 was established from a poorly differentiated esophageal squamous cell carcinoma and is widely used in esophageal cancer research. The adherent epithelial cells retain certain barrier characteristics of the native tissue while displaying hallmarks of malignancy, including dysregulated proliferation, migration, and invasion. Their aggressive phenotype and genetic background render them particularly suitable for exploring the role of epigenetic modifiers in tumor progression and therapeutic resistance.
DNMT3A is a de novo DNA methyltransferase that catalyzes the transfer of methyl groups from S-adenosyl methionine (SAM) to CpG dinucleotides, leading to chromatin compaction and transcriptional repression. It is regulated upstream by SP1 transcription factor and RAS signaling, and post-transcriptionally repressed by the miR-29 microRNA family. DNMT3A interacts with DNMT3L, which enhances its activity, as well as with UHRF1, HDAC1, and chromatin remodeling complexes to methylate and silence tumor suppressor genes such as CDKN2A, RASSF1A, and HOXA clusters. By integrating oncogenic signals, DNMT3A enforces an epigenetic landscape that promotes tumorigenesis.
In KYSE-150 cells, DNMT3A knockout abolishes de novo methylation activity, potentially leading to reactivation of aberrantly silenced tumor suppressors and reversal of malignant epigenetic programs. The loss of DNMT3A is expected to cause genome-wide demethylation, especially at promoter CpG islands, and may render cells more susceptible to treatment with hypomethylating agents such as decitabine or azacitidine. The polyclonal nature of this model minimizes clonal bias, yielding more reliable and reproducible phenotypes for studying epigenetic addiction in esophageal squamous cell carcinoma.
These polyclonal knockout cells are suited for bisulfite sequencing, methylated DNA immunoprecipitation, and ChIP-qPCR to profile DNA methylation and chromatin changes. RNA-seq enables transcriptome-wide assessment of gene reactivation, while functional assays such as proliferation, migration, and invasion quantify phenotypic consequences. Additionally, they serve as a platform for screening hypomethylating agents and other compounds that synergize with DNMT3A loss. For further information, please contact Ascent Research.
