The CHD3 Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the CHD3 gene in the human Raji B lymphocyte line. This heterogeneous pool contains a spectrum of loss-of-function mutations resulting from CRISPR/Cas9-mediated gene disruption, preserving the inherent genetic diversity of the edited population and avoiding clonal selection biases. The polyclonal format provides a robust and reproducible model for interrogating NuRD complex-dependent processes without the confounding effects of monoclonal expansion.
Raji cells are an EBV-positive Burkitt lymphoma-derived B lymphocyte line that exhibits hallmarks of mature B cells, including surface immunoglobulin expression, antigen presentation, and antibody secretion. Their transformed, yet lineage-committed, phenotype makes them a widely utilized model for studying B-cell malignancies, lymphoma biology, and immune cell function. The presence of the EBV genome further facilitates investigations into viral-driven oncogenesis and its interplay with host epigenetic machinery.
CHD3 encodes the chromodomain-helicase-DNA-binding protein 3, the central ATPase subunit of the nucleosome remodeling and deacetylase (NuRD) complex. Within this multi-subunit assembly, CHD3 directly interacts with scaffold proteins MTA1 and MTA2, histone deacetylases HDAC1 and HDAC2, histone-binding proteins RBBP4 and RBBP7, methyl-CpG binding proteins MBD2 and MBD3, and the co-repressors GATAD2A and GATAD2B. Recruitment of NuRD to chromatin is mediated by MBD2/3 binding to methylated DNA, while upstream kinase CDK2 phosphorylates CHD3 to modulate its activity. Once targeted, CHD3 hydrolyzes ATP to slide nucleosomes, and this remodeling is functionally coupled to HDAC-mediated histone deacetylation, resulting in robust transcriptional repression of downstream genes. Key transcriptional targets include cell cycle inhibitors, linking CHD3 activity to the regulation of proliferation and survival.
Disruption of CHD3 in Raji B cells abrogates the core enzymatic activity of the NuRD complex, impairing the coordination between ATP-dependent chromatin remodeling and histone deacetylation. This loss-of-function leads to aberrant chromatin states and derepression of NuRD target genes, potentially perturbing cell cycle progression, apoptotic signaling, and differentiation pathways. Given the dependency of Burkitt lymphoma cells on epigenetic regulation for sustaining oncogenic transcriptional programs, this knockout model offers a physiologically relevant system to dissect the role of NuRD in B-cell transformation. Moreover, mutations in CHD3 are associated with neurodevelopmental disorders, underscoring the broader biological significance of this chromatin remodeler.
Researchers can leverage these polyclonal CHD3 knockout Raji cells for diverse applications, including mechanistic studies of NuRD complex function in B lymphocytes, high-content screening for small molecules that modulate chromatin remodeling, and validation of CHD3 as a therapeutic target for hematological malignancies. Compatible downstream assays include RT-qPCR and RNA-seq for transcriptome analysis, ChIP-qPCR for profiling histone modification changes, co-immunoprecipitation to assess NuRD complex integrity, and flow cytometry combined with immunofluorescence for cell cycle and apoptosis phenotyping. Drug sensitivity assays can further elucidate the impact of CHD3 loss on therapeutic responses. For additional information, please contact Ascent Research.
