The DNMT3A Knockout T-47D Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population with disrupted DNMT3A gene function. This heterogeneous pool of T-47D cells harbors a variety of loss-of-function alleles, eliminating the need for single-cell cloning and preserving population-level genetic diversity. The product provides a robust model for studying DNMT3A deficiency without clonal artifacts.
T-47D is a human mammary epithelial carcinoma cell line isolated from a pleural effusion of a ductal carcinoma. It is estrogen receptor-positive and expresses progesterone and androgen receptors, making it a widely used model for hormone-responsive luminal A breast cancer. T-47D cells retain key features of hormone-dependent growth and are ideal for epigenetic research in breast cancer.
DNMT3A encodes a de novo DNA methyltransferase that establishes DNA methylation patterns by adding methyl groups to unmethylated CpG sites, resulting in transcriptional silencing. The enzyme is regulated by upstream factors such as E2F1, STAT3, PI3K/AKT, and RAS/MAPK signaling, and by the miR-29 microRNA family. DNMT3A interacts with DNMT3L, HDAC1, EZH2, UHRF1, and PCNA within epigenetic complexes. It collaborates with the maintenance methyltransferase DNMT1 to ensure methylation inheritance, and its activity is recognized by MBD proteins that promote chromatin compaction. Its methylation activity directly represses tumor suppressors including CDKN2A, CDH1, RASSF1A, and MLH1, contributing to breast cancer pathogenesis.
In T-47D cells, DNMT3A knockout disrupts the DNA methylation landscape, potentially reactivating silenced tumor suppressor genes and altering hormone-responsive phenotypes. This model allows dissection of DNMT3A-specific contributions to breast cancer epigenetics, particularly in the context of PI3K/AKT pathway crosstalk. The polyclonal nature preserves functional heterogeneity, reflecting the diversity of methylation patterns observed in tumors.
Applications include epigenetic gene regulation studies, screening of hypomethylating agents such as decitabine, and genome-wide methylation analysis by bisulfite sequencing. Researchers can compare transcriptional profiles using RNA-seq and RT-qPCR, map chromatin changes via ChIP, and assess functional consequences with cell proliferation and drug sensitivity assays. Western blotting and RT-qPCR are recommended for confirming DNMT3A disruption. For ordering and technical inquiries, contact Ascent Research.
