The DNMT3A Knockout A-549 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population originating from the A-549 human lung adenocarcinoma cell line. This product provides a loss-of-function model for DNMT3A, a de novo DNA methyltransferase responsible for establishing CpG methylation patterns during development. CRISPR/Cas9-mediated gene disruption yields a heterogeneous pool of edited cells, reflecting the natural variability of knockout outcomes without clonal selection, and is ideal for functional studies of epigenetic regulation.
A-549 cells are derived from alveolar basal epithelial cells and represent a widely used non-small cell lung cancer (NSCLC) model. They maintain alveolar epithelial characteristics, including barrier and secretory functions, and harbor genomic alterations that drive tumorigenesis. The cell line exhibits aberrant DNA methylation, making it particularly relevant for investigating DNMT3A-dependent epigenetic silencing in a lung adenocarcinoma context.
DNMT3A catalyzes de novo CpG methylation using S-adenosyl methionine (SAM), with patterns maintained by DNMT1. Its expression is regulated by SP1, IL-6/STAT3, WNT/??-catenin, and TGF-??. DNMT3A complexes with DNMT3L and interacts with EZH2 (PRC2), HDAC1/2, UHRF1, and MYC. It targets methylation of tumor suppressors CDKN2A, RASSF1A, the HOXA cluster, and facilitates TP53 silencing. Knockout disrupts these interactions, derepressing silenced genes and altering chromatin states.
In A-549 cells, DNMT3A knockout induces promoter hypomethylation, potentially reactivating tumor suppressors and influencing cell differentiation and proliferation. This polyclonal model captures the heterogeneity of epigenetic responses, providing a physiologically relevant system to study DNA methylation dysregulation found in lung adenocarcinoma and related malignancies such as acute myeloid leukemia and myelodysplastic syndromes.
Applications include bisulfite sequencing for methylation analysis, RNA-seq and RT-qPCR for expression profiling, ChIP-qPCR for histone marks, and functional assays like proliferation, apoptosis, and drug sensitivity screening with decitabine. The model supports tumor suppressor reactivation and differentiation studies. For further details, contact Ascent Research.
