The DNMT3A Knockout LoVo Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal population in which the human DNMT3A gene has been disrupted. Derived from the LoVo colorectal adenocarcinoma line, these polyclonal knockout cells provide a loss-of-function model to study de novo DNA methylation in cancer epigenetics, gene silencing, and tumor suppression. The pooled format, free from single-cell cloning, captures diverse editing outcomes while supporting reproducible functional studies. Validated population-level gene disruption enables investigation of DNMT3A-dependent mechanisms in colorectal cancer biology.
The LoVo cell line, established from a lymph node metastasis of a 56-year-old male with Dukes?? type C colorectal adenocarcinoma, is an adherent epithelial model widely used in colorectal cancer research. This line carries mutations in key oncogenic pathways, including Wnt/??-catenin and p53, and exhibits aberrant DNA methylation patterns characteristic of colorectal tumors. Its metastatic origin and genetic profile make it a valuable system for dissecting the interplay between genetic alterations and epigenetic dysregulation in colorectal carcinogenesis.
DNMT3A is a de novo DNA methyltransferase that methylates CpG sites to establish epigenetic silencing. It interacts with DNMT3L, HDAC1/2, EZH2, SUZ12, and UHRF1, and is regulated by TP53, SP1/SP3, MAPK/PI3K/AKT signaling, and S-adenosyl methionine levels. Its targets include promoters of tumor suppressors such as CDKN2A, MLH1, APC, PTEN, SFRP1, and DKK1, leading to recruitment of methyl-binding proteins, histone deacetylation, and gene repression. This functionally connects DNMT3A to Wnt/??-catenin signaling via interaction with ??-catenin (CTNNB1).
In LoVo cells, DNMT3A knockout disrupts the aberrant DNA methylation responsible for silencing tumor suppressor genes, potentially reactivating their expression and restoring growth-suppressive signaling. This model enables dissection of DNMT3A-specific contributions to the colorectal cancer epigenome, distinct from other methyltransferases, and clarifies its role downstream of oncogenic pathways. The knockout thus provides a precise tool to probe causal relationships between de novo methylation and cancer phenotypes such as uncontrolled proliferation, evasion of apoptosis, and metastatic potential.
This polyclonal knockout cell pool supports a wide range of epigenetic applications: screening DNA hypomethylating agents (e.g., 5-azacytidine, decitabine), analyzing promoter methylation by bisulfite sequencing or methylation-specific PCR, and assessing transcriptome-wide changes via RNA-seq. Protein-level validation is performed by western blotting and RT-qPCR, while chromatin occupancy studies use ChIP-qPCR. Functional outcomes are measured through proliferation, colony formation, apoptosis, migration, and invasion assays, facilitating both fundamental epigenetic research and preclinical drug discovery. For technical inquiries, please contact Ascent Research.
