The DNMT3B Knockout HAP1 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human HAP1 near-haploid chronic myeloid leukemia cell line, designed for loss-of-function studies of the DNMT3B gene. This heterogeneous knockout model targets the gene encoding the de novo DNA methyltransferase DNMT3B, providing a versatile tool to investigate epigenetic regulation mechanisms in a genetically tractable background.
HAP1 is a well-characterized near-haploid human cell line established from a male patient with chronic myeloid leukemia. Its near-haploid karyotype simplifies genetic manipulation and knockout generation, making it a preferred host for CRISPR-based functional genomics. The line retains key leukemic features and supports reproducible cell-based assays, including proliferation, colony formation, and drug sensitivity testing.
DNMT3B is a pivotal de novo DNA methyltransferase that establishes DNA methylation patterns during development by catalyzing the transfer of methyl groups to unmethylated CpG dinucleotides. It functions within a complex network: transcriptionally activated by SP1 and downstream of STAT3 signaling triggered by IL-6, while being post-transcriptionally repressed by the miR-29 family. DNMT3B physically interacts with maintenance methyltransferase DNMT1, its paralog DNMT3A, heterochromatin protein HP1, histone deacetylases HDAC1/2, ubiquitin ligase UHRF1, and the replication factor PCNA. These interactions coordinate DNA methylation with histone modifications and chromatin remodeling. Key downstream targets silenced by DNMT3B-mediated methylation include the cell cycle inhibitor CDKN2A (p16), the adhesion molecule CDH1 (E-cadherin), and the tumor suppressor RASSF1A, along with repetitive retrotransposon sequences, thereby linking DNMT3B to control of proliferation, invasion, and genomic integrity.
In the HAP1 leukemia model, ablation of DNMT3B disrupts aberrant methylation patterns associated with oncogenesis. This knockout population permits dissection of DNMT3B??s role in silencing tumor suppressor genes, enabling researchers to examine how loss of methylation reactivates pathways that restrain leukemic growth. The near-haploid background reduces genetic redundancy, allowing clearer phenotypic attribution to DNMT3B in epigenetic drug resistance and chromatin state analyses.
Applications for this polyclonal knockout model are broad, including bisulfite sequencing to map global methylation changes, methylation-specific PCR for candidate gene validation, RNA-seq to profile transcriptomic alterations, and western blotting to confirm target protein re-expression. The cells are also suitable for ChIP-qPCR of histone marks (e.g., H3K9me3, H3K27me3) to study crosstalk between DNA methylation and histone modifications, and for functional assays such as cell proliferation, colony formation, and sensitivity profiling to hypomethylating agents like decitabine. This product is an ideal resource for cancer epigenetics, ICF syndrome modeling, and drug discovery programs. For additional information, please contact Ascent Research.
