The DNMT3A Knockout HGC-27 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population in which the DNMT3A gene has been disrupted in the human gastric carcinoma cell line HGC-27. This heterogeneous pool of knockout cells provides a robust loss-of-function model for interrogating DNMT3A-dependent epigenetic control mechanisms in a cancer-relevant background, without the bias of clonal selection.
HGC-27 is an epithelial cell line originating from a metastatic lymph node of a gastric adenocarcinoma, making it a widely accepted model for gastric cancer metastasis studies and drug screening. Its metastatic derivation renders it particularly valuable for dissecting the molecular underpinnings of tumor cell dissemination and for evaluating therapeutic interventions targeting metastatic progression.
DNMT3A encodes a de novo DNA methyltransferase responsible for establishing and maintaining DNA methylation patterns, primarily through the transfer of methyl groups to cytosine residues in CpG dinucleotides, thereby mediating transcriptional silencing. The activity and expression of DNMT3A are regulated by multiple upstream signals, including the transcription factors E2F1, OCT4, and SOX2, as well as the Wnt/beta-catenin and MAPK/ERK pathways, with post-transcriptional modulation by the miR-29 family. Functionally, DNMT3A interacts with DNMT3L, HDAC1, HDAC2, UHRF1, and PCNA, and collaborates with H3K4me0 and H3K36me3 histone modifications to direct methylation. Its silencing targets encompass critical tumor suppressors such as CDKN2A/p16, CDKN2B/p15, RASSF1A, and MLH1, as well as stemness-associated genes OCT4 and NANOG, downstream of DNA methylation readers like MBD proteins and HDACs.
In the HGC-27 background, disruption of DNMT3A leads to the loss of de novo methylation and consequent reactivation of silenced tumor suppressor loci, including CDKN2A and RASSF1A. This derepression is anticipated to impair proliferation, migratory capacity, and metastatic potential, thereby modeling key aspects of epigenetic dysregulation observed in gastric adenocarcinoma and offering a platform to study the role of DNA methylation in tumor maintenance and metastasis.
This knockout cell population is optimally suited for a variety of advanced research applications, including epigenetic cancer biology studies, genome-wide DNA methylation analysis via bisulfite sequencing or RNA-seq, drug sensitivity profiling for DNMT inhibitors and other epigenetic therapies using MTT assays, and functional investigations of metastasis through migration/invasion and colony formation assays. Additionally, it enables the biochemical verification of target gene reactivation by western blotting and RT-qPCR, as well as chromatin-level analysis via ChIP-qPCR. For further technical details or to request a quotation, please contact Ascent Research.
