The DNMT3A Knockout KYSE-30 Polyclonal Cells product consists of a CRISPR/Cas9-edited polyclonal knockout cell population with targeted disruption of the DNMT3A gene in the human KYSE-30 esophageal squamous cell carcinoma line. This loss-of-function model is generated via CRISPR/Cas9-mediated gene disruption, resulting in a heterogeneous pool of edited cells that serves as a powerful tool for investigating de novo DNA methylation and epigenetic regulation without the need for single-cell cloning.
KYSE-30 is a well-differentiated human esophageal squamous cell carcinoma line derived from a primary tumor, providing a physiologically relevant model for esophageal cancer research. These cells retain key genomic and signaling characteristics of esophageal SCC, making them an appropriate host for studying the epigenetic mechanisms that contribute to malignancy.
DNMT3A encodes a de novo DNA methyltransferase responsible for establishing DNA methylation patterns during development. Its expression is regulated by transcription factors SP1 and STAT3, as well as RAS/ERK signaling, and is targeted by the miR-29 microRNA family. DNMT3A interacts with DNMT3L, DNMT1, UHRF1, and histone modifiers HDAC1/2 and EZH2, linking DNA methylation to repressive chromatin. It is recruited to H3K36me3-marked regions and methylates promoters of tumor suppressor genes such as CDKN2A (p16) and CDH1 (E-cadherin), leading to their transcriptional silencing.
Disruption of DNMT3A in KYSE-30 cells abolishes de novo methylation, causing promoter hypomethylation and reactivation of silenced tumor suppressors. This epigenetic reprogramming alters gene expression programs involved in proliferation, apoptosis, and differentiation, thereby impacting esophageal cancer cell behavior. As DNMT3A mutations are implicated in clonal hematopoiesis and hematologic malignancies, this model also enables comparative studies of epigenetic mechanisms across tumor types.
This polyclonal knockout population is suited for DNA methylation analysis using bisulfite sequencing and methylation-specific PCR, gene expression profiling via RNA-seq and RT-qPCR, and chromatin studies with ChIP-qPCR. Functional assays such as proliferation and apoptosis measurements, western blotting, and drug screening facilitate the exploration of DNMT3A-dependent phenotypes and therapeutic reactivation strategies. For more information or a quote, please contact Ascent Research.
