The DNMT3A Knockout 786-O Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population of human 786-O renal cell adenocarcinoma cells. This loss-of-function model results from disruption of the DNMT3A gene, producing a heterogeneous pool of edited alleles that reflects natural genetic variation and avoids clonal artifacts. The knockout enables stable ablation of DNMT3A protein expression, making it a robust system for epigenetic studies in a VHL-mutant kidney cancer context.
The 786-O line is a widely used epithelial model derived from a clear cell renal cell carcinoma (ccRCC) tumor. It carries a biallelic VHL inactivation, leading to constitutive stabilization of hypoxia-inducible factors (HIFs) and chronic activation of hypoxic gene programs under normoxia. This background recapitulates the hallmark molecular feature of sporadic ccRCC and provides a relevant platform for investigating the interplay between DNA methylation and HIF-driven oncogenesis. The cells display aggressive properties including high proliferation, migration, and invasion, which are amenable to functional dissection in knockout models.
DNMT3A is a de novo DNA methyltransferase that transfers methyl groups from S-adenosyl methionine (SAM) to CpG dinucleotides, establishing methylation patterns for gene silencing. Its activity is regulated by STAT3, NF-??B, MYC, and TGF-?? signaling. DNMT3A complexes with DNMT3L, HDAC1/2, EZH2, UHRF1, and PCNA to mediate epigenetic repression. Key downstream targets include tumor suppressor promoters CDKN2A, CDKN1A, RASSF1A, and APC. Disruption of DNMT3A leads to genome-wide hypomethylation and potential reactivation of these silenced loci.
In the VHL-mutant 786-O background, DNMT3A knockout is expected to abolish de novo methylation capability, resulting in global DNA hypomethylation and transcriptional derepression of hypermethylated tumor suppressor genes. This epigenetic rewiring may either augment or attenuate HIF-mediated oncogenic signals, offering a unique system to study crosstalk between methylation and hypoxia pathways in renal carcinogenesis. The model can be used to examine how loss of methylation influences cell proliferation, migration, drug sensitivity, and tumor suppressor expression, particularly in response to demethylating agents such as decitabine and azacitidine.
The DNMT3A Knockout 786-O Polyclonal Cells are suited for a broad spectrum of epigenetic and cancer biology assays. Whole-genome or reduced representation bisulfite sequencing allows comprehensive DNA methylation profiling, while RNA-seq and RT-qPCR quantify transcriptomic changes. Functional readouts include colony formation, wound healing, Transwell invasion, and cell viability assays following treatment with DNA methyltransferase inhibitors (azacitidine, decitabine). Chromatin modifications can be assessed by ChIP-qPCR for histone marks and immunofluorescence for 5-methylcytosine. This product is delivered as a live polyclonal population, enabling direct culture and expansion. For technical inquiries, pricing, or protocol support, reach out to Ascent Research.
