DNMT3A Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population of HT29 cells harboring targeted disruption of the DNMT3A gene. This knockout model provides a powerful tool for investigating de novo DNA methylation mechanisms, epigenetic regulation, and colorectal cancer biology. The polyclonal format reflects a heterogeneous knockout pool generated without single-cell cloning, enabling immediate use in functional genomics and drug screening applications.
HT29 is a human colorectal adenocarcinoma epithelial cell line derived from a primary tumor of a female patient. It exhibits intestinal differentiation markers and is widely employed as a model for intestinal epithelial barrier function and colorectal cancer. The cells retain key signaling pathways relevant to colorectal carcinogenesis, including Wnt/??-catenin and TP53 networks, making them particularly suitable for dissecting epigenetic alterations in tumorigenesis.
DNMT3A encodes a de novo DNA methyltransferase that catalyzes the transfer of methyl groups from S-adenosyl methionine (SAM) to cytosine residues in CpG dinucleotides, establishing DNA methylation patterns critical for gene regulation and genomic stability. DNMT3A is regulated by transcription factors SP1 and SP3, retinoic acid receptor signaling, and the Wnt/??-catenin pathway, while it functionally interacts with DNMT3L, UHRF1, PCNA, EZH2, HP1 proteins, and histone H3K36me3. Its methyltransferase activity leads to CpG island hypermethylation of tumor suppressor genes such as CDKN2A, MLH1, and APC, contributing to transcriptional repression and chromatin remodeling.
In the context of HT29 cells, DNMT3A loss disrupts de novo methylation processes, potentially reactivating silenced tumor suppressors and altering chromatin structure. This knockout model enables researchers to dissect the causal role of DNA methylation in colorectal cancer cell proliferation, differentiation, and drug response. It is particularly valuable for investigating the interplay between epigenetic silencing and oncogenic signaling pathways in a well-characterized colorectal adenocarcinoma background.
Typical applications include epigenetic regulation studies, global DNA methylation quantification by LC-MS, bisulfite sequencing to map methylation changes, and transcriptome profiling via RNA-seq to identify reactivated genes. Functional assays such as colony formation and drug sensitivity testing with demethylating agents (e.g., 5-azacytidine) are also routinely employed. Additionally, the cells can be used for chromatin immunoprecipitation (ChIP-qPCR) to assess histone modification changes. For further information on this knockout model, please contact Ascent Research.
