DNMT3A Knockout Ca Ski Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population generated from the Ca Ski human cervical carcinoma line. This product provides a heterogeneous loss-of-function model for studying DNMT3A-mediated epigenetic regulation in cancer, with target-gene disruption achieved across the population without clonal selection.
The Ca Ski cell line was derived from a metastatic cervical epidermoid carcinoma and harbors integrated HPV-16 genomes, making it a key model for HPV-positive cervical cancer research. These epithelial cells retain expression of viral oncoproteins E6 and E7, which target p53 and Rb, and serve as a relevant system for exploring epigenetic disruptions linked to viral carcinogenesis.
DNMT3A is a de novo DNA methyltransferase that catalyzes the methylation of unmethylated CpG dinucleotides, contributing to gene silencing and chromatin remodeling. Its expression is regulated by upstream factors including the transcription factor SP1, STAT3, the miR-29 microRNA family, and HIF1??. DNMT3A forms complexes with DNMT3L, HDAC1, HP1, UHRF1, and PCNA, integrating DNA methylation with histone deacetylation and replication-coupled maintenance. Downstream targets include tumor suppressor genes such as CDKN2A (p16), CDH1 (E-cadherin), and RASSF1, whose transcriptional repression is mediated by promoter hypermethylation. The broader pathway involves S-adenosyl methionine as a methyl donor, methyl-CpG-binding domain proteins (MBDs) as methylation readers, and TET enzymes as demethylases.
In the context of HPV-16-positive Ca Ski cells, aberrant DNA methylation driven by DNMT3A synergizes with viral oncoproteins to silence critical tumor suppressors, promoting cervical carcinogenesis. Knockout of DNMT3A in this background is anticipated to reverse methylation-dependent silencing of genes like CDH1 and CDKN2A, restore their expression, and suppress malignant phenotypes such as enhanced proliferation and invasion. This polyclonal model recapitulates tumor heterogeneity more faithfully than clonal lines and offers a powerful tool to dissect the interplay between viral infection and epigenetic reprogramming.
These polyclonal knockout cells support a wide range of epigenetics applications, including genome-wide methylation analysis by bisulfite sequencing, transcriptome profiling via RNA-seq, and targeted gene expression analysis by RT-qPCR. Chromatin immunoprecipitation (ChIP) can assess histone modification changes, while functional assays such as proliferation, migration, and invasion quantify tumorigenic capacity. The model is particularly suited for drug screening with demethylating agents like 5-azacytidine. For additional product information or custom requests, please contact Ascent Research.
