The DNMT3A Knockout UM-UC-3 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population disrupting the DNMT3A gene in the human bladder transitional cell carcinoma cell line UM-UC-3. This polyclonal product provides a robust loss-of-function model, avoiding clonal selection biases and presenting a heterogeneous mixture of edited cells that collectively ablate DNMT3A function. The knockout model is a valuable tool for investigating the role of de novo DNA methylation in cancer epigenetics and tumorigenesis.
The host cell line UM-UC-3 originates from a male bladder transitional cell carcinoma, representing a malignant urothelial cell model widely utilized in bladder cancer research. UM-UC-3 cells exhibit characteristics of high-grade bladder carcinoma, including aberrant proliferative signaling and metastatic potential. This cell line is particularly valuable for studying the molecular underpinnings of urothelial carcinogenesis, drug response, and epigenetic dysregulation in solid tumors.
DNMT3A is a de novo DNA methyltransferase that establishes DNA methylation patterns by catalyzing the transfer of methyl groups from S-adenosyl methionine (SAM) to cytosine residues within CpG dinucleotides. Its activity is regulated by upstream signals including PI3K/AKT signaling, mitogenic stimuli, and transcription factors PU.1 (SPI1) and GATA1. DNMT3A functions within multiprotein complexes containing DNMT3L, UHRF1, HDAC1, PCNA, and EZH2, and cooperates with nucleosome remodeling factors. This enzyme transcriptionally represses key tumor suppressor genes, including CDKN2A (p16), MLH1, BRCA1, and RB1, via promoter methylation, leading to epigenetic gene silencing. Additionally, DNMT3A integrates with the Wnt/??-catenin pathway to influence cell fate. Methylation marks are recognized by methyl-CpG-binding domain (MBD) proteins and MeCP2, which propagate chromatin compaction and stable gene repression.
In the context of UM-UC-3 bladder carcinoma cells, DNMT3A knockout eliminates the primary de novo methylation activity, potentially reactivating silenced tumor suppressor genes and reversing malignant epigenetic states. This disruption may impair cancer cell proliferation, enhance sensitivity to therapeutic agents, and provide a unique platform to dissect the interplay between DNA methylation and oncogenic signaling pathways. The model is particularly relevant for studying how epigenetic silencing contributes to bladder cancer progression, metastasis, and chemoresistance.
Applications include epigenetics research, functional genomics, cancer biology, and drug target validation. Researchers can perform bisulfite sequencing to assess DNA methylation changes, RT-qPCR and western blotting to confirm knockout and derepression of targets such as CDKN2A, and cell proliferation or colony formation assays to evaluate phenotypic effects. The model is also suitable for drug sensitivity screens to identify epigenetic therapies. For additional information, please contact Ascent Research.
