DNMT3A Knockout HEK293T Polyclonal Cells comprise a CRISPR/Cas9-edited polyclonal knockout cell population derived from HEK293T, providing constitutive disruption of the DNMT3A gene. This heterogeneous loss-of-function model enables study of de novo DNA methylation without clonal selection bias. Targeted gene disruption abolishes DNMT3A methyltransferase activity, allowing investigation of its immediate and downstream epigenetic effects in a widely used human cell background.
The host HEK293T line originates from human embryonic kidney cells and stably expresses the SV40 large T-antigen, which drives episomal plasmid replication. This yields high recombinant protein expression and efficient lentivirus production. HEK293T cells are valued for robust growth, high transfection efficiency, and broad biochemical utility, making the DNMT3A knockout derivative a reliable platform for epigenetic research.
DNMT3A functions as a de novo DNA methyltransferase that establishes methylation patterns by transferring a methyl group from S-adenosylmethionine to cytosine in CpG dinucleotides, generating 5-methylcytosine and inducing transcriptional silencing. Its activity is controlled by upstream transcription factors OCT4, SOX2, MYC, and E2F1, and it operates within a complex including DNMT3B, DNMT3L, HDAC1, HDAC2, EZH2, HP1, and UHRF1. This machinery promotes silencing of tumor suppressor genes such as CDKN2A, CDKN2B, RASSF1A, and MLH1. In the knockout cells, loss of DNMT3A disrupts these interactions, leading to global hypomethylation and derepression of methylation-sensitive loci, thus providing a clean background to dissect how DNMT3A integrates signals from pluripotency factors and chromatin regulators.
In HEK293T cells, DNMT3A knockout removes the major de novo methylation activity, preventing methylation-dependent silencing of introduced transgenes and viral promoters. This polyclonal population sustains expression of methylation-sensitive constructs, facilitating epigenetic studies without confounding DNMT3A activity. The SV40 T-antigen machinery remains intact, retaining capability for viral packaging and protein production. This model allows examination of the interplay between replication, chromatin, and DNA methylation in a defined hypomethylated context.
These cells are applicable to cancer epigenetics research, including acute myeloid leukemia, myelodysplastic syndromes, and Tatton-Brown?CRahman overgrowth syndrome. They support drug sensitivity assays with hypomethylating agents such as decitabine and enable DNA methyltransferase inhibitor screening. The knockout model is suitable for stem cell differentiation studies and epigenetic reprogramming experiments. Representative assays include Western blotting, RT-qPCR for re-expression of methylation-silenced genes (e.g., CDKN2A, RASSF1A), bisulfite sequencing, MeDIP, and ChIP-qPCR for histone modifications. For detailed technical inquiries, contact Ascent Research.
