The DNMT3B Knockout HCT 116 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population targeting the DNMT3B gene in the human HCT 116 colorectal carcinoma cell line. This loss-of-function model is generated by CRISPR/Cas9-mediated disruption of the DNMT3B locus, producing a heterogeneous pool of cells with targeted gene ablation. The polyclonal format retains population diversity, suitable for studying DNMT3B-dependent phenotypes without the biases of clonal selection. As a polyclonal knockout pool, it enables robust analysis of DNA methylation dynamics and epigenetic gene silencing in a cancer-relevant background.
HCT 116 is a widely used human colorectal carcinoma epithelial cell line, isolated from a male patient. It harbors a KRAS G13D mutation and is MLH1-deficient, leading to high microsatellite instability (MSI-H). These genetic features make it a valuable model for colorectal cancer research, particularly for epigenetic regulation, mismatch repair deficiency, and oncogenic signaling. The adherent growth and well-characterized genome facilitate reproducible experiments and integration into diverse assays.
DNMT3B encodes a de novo DNA methyltransferase that establishes methylation patterns at CpG islands and repetitive elements. It is regulated by upstream factors such as MYC, E2F1, SP1, and STAT3, and post-transcriptionally repressed by the miR-29 family. DNMT3B interacts with DNMT1, DNMT3L, HDAC1/2, PCNA, UHRF1, and EZH2 to coordinate methylation and chromatin remodeling. Using S-adenosyl methionine (SAM) as a methyl donor, it produces 5-methylcytosine, which recruits MBD2 and MeCP2 to repress transcription. Key downstream targets include tumor suppressor gene promoters (CDKN2A, RASSF1A, SOCS1) and repetitive elements (LINE-1, pericentromeric repeats).
In HCT 116 cells, DNMT3B knockout reduces global DNA methylation, reactivating silenced tumor suppressors and impairing proliferation. The polyclonal population enables assessment of heterogeneous methylation changes without clonal bias. Given the MLH1-deficient, MSI-H background, this model is ideal for studying the interplay between DNA methylation and mismatch repair, and for investigating tumorigenic potential and cell cycle regulation.
Applications include epigenetic studies (bisulfite sequencing, RNA-seq), drug screening for demethylating agents, and tumor suppressor reactivation assays. The cells are also suitable for colony formation, flow cytometry, and xenograft models. They support chromatin analysis and gene expression profiling to map DNMT3B targets and interactors. For further information, please contact Ascent Research.
