DNMT3A Knockout 143B Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human osteosarcoma cell line 143B, featuring targeted disruption of the de novo DNA methyltransferase DNMT3A. This polyclonal format provides a heterogeneous pool of knockout genotypes, avoiding artifacts associated with single-cell cloning while enabling robust loss-of-function studies. The product is designed for researchers investigating epigenetic regulation in cancer, particularly the role of DNA methylation in gene silencing and tumor suppressor reactivation.
The 143B cell line is a highly tumorigenic and metastatic human osteosarcoma model, retaining aggressive growth and metastatic potential characteristic of bone cancer. Widely used in oncology research, 143B cells are well-suited for examining tumor biology, metastasis, and epigenetic mechanisms driving malignancy. Their established in vitro and in vivo tractability makes them an appropriate host for dissecting DNMT3A-dependent processes in a disease-relevant context.
DNMT3A catalyzes de novo DNA methylation, establishing CpG methylation patterns that mediate epigenetic silencing. It is transcriptionally regulated by factors including SP1, NF-??B, and STAT3, with upstream modulation by IL-6 and Wnt signaling. DNMT3A forms functional complexes with DNMT3L, HDAC1/2, EZH2, and UHRF1, coordinating chromatin repression. Disruption of DNMT3A leads to hypomethylation of target gene promoters, such as CDKN2A, CDKN2B, RASSF1A, and p14ARF, potentially reactivating these tumor suppressors and altering downstream signaling.
In the 143B osteosarcoma background, DNMT3A knockout abrogates de novo methylation capacity, likely reversing epigenetic silencing of key tumor suppressors and attenuating malignant phenotypes. The model captures population-level epigenetic heterogeneity through its polyclonal nature, enabling studies of DNA methylation dynamics without clonal bias. It provides a platform to explore connections between DNMT3A loss, hypomethylation, and pathways relevant to osteosarcoma progression and clonal hematopoiesis.
Typical applications include bisulfite sequencing to map methylation changes, RT-qPCR or RNA-seq for gene expression profiling, and Western blotting to confirm tumor suppressor re-expression. Functional assays??such as cell proliferation, colony formation, and migration??can assess the impact of DNMT3A disruption on osteosarcoma cell behavior. This knockout model supports investigation of epigenetic therapies, DNMT inhibitor responses, and Wnt pathway crosstalk. For further information or custom requests, please contact Ascent Research.
