The DNMT3A Knockout PaTu 8988t Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed for loss-of-function studies of the DNA methyltransferase 3 alpha (DNMT3A) gene. Through CRISPR/Cas9-mediated gene disruption, this product abolishes DNMT3A protein expression, providing a reliable model to investigate the consequences of DNMT3A deficiency in a human pancreatic cancer background. The polyclonal format ensures representation of a diverse array of editing events, offering a robust population-level knockout effect suitable for epigenomic and functional assays.
The parental PaTu 8988t cell line is a human pancreatic ductal adenocarcinoma (PDAC) model harboring an activating KRAS G12V mutation, a hallmark driver of pancreatic tumorigenesis. This cell line is widely employed to study PDAC biology, including signaling networks, drug response, and metastatic mechanisms. Its well-characterized genetic background makes it an ideal host for creating gene knockouts to dissect oncogenic and tumor-suppressive pathways in a disease-relevant context.
DNMT3A encodes a de novo DNA methyltransferase that catalyzes the transfer of methyl groups to CpG dinucleotides, establishing DNA methylation patterns critical for epigenetic regulation of gene expression and chromatin remodeling. DNMT3A is activated by upstream regulators such as SP1, STAT3, and PU.1, and functions in concert with interacting partners including DNMT3L, DNMT1, HDAC1, and EZH2. Its enzymatic activity targets tumor suppressor genes (e.g., CDKN2A, MLH1), repetitive elements, and imprinted loci, leading to transcriptional silencing. The knockout disrupts this de novo methylation machinery, resulting in genome-wide hypomethylation and reactivation of aberrantly silenced genes.
In the context of PaTu 8988t cells, DNMT3A knockout holds particular significance for pancreatic cancer research. PDAC is characterized by extensive epigenetic alterations, including aberrant CpG island methylation that silences key tumor suppressors. By eliminating DNMT3A activity in a KRAS mutant background, these polyclonal knockout cells enable the study of how loss of de novo methylation influences pancreatic cancer cell behavior, potentially uncovering vulnerabilities associated with epigenetic dysregulation. This model is valuable for exploring the interplay between KRAS-driven signaling and DNA methylation, and for identifying synthetic lethal interactions with methylation inhibitors.
Typical research applications include cancer epigenetics, tumor suppressor gene reactivation studies, and DNA methylation inhibitor screening. The knockout cells are suitable for assays such as western blotting and RT-qPCR to confirm DNMT3A ablation and target gene expression changes, bisulfite sequencing and methylated DNA immunoprecipitation (MeDIP) to map methylation alterations, and cell proliferation and drug sensitivity assays to assess functional consequences. For further information or to discuss custom applications, please contact Ascent Research.
