The H2BC12 Knockout HeLa Polyclonal Cells represent a CRISPR/Cas9-mediated gene-disrupted polyclonal cell population targeting the human H2BC12 locus in the widely utilized HeLa cell line. This product provides researchers with a heterogeneous pool of edited cells harboring targeted disruptions of the H2BC12 gene, enabling loss-of-function studies without single-cell clonal selection. The polyclonal nature preserves genetic diversity and mitigates clonal artifacts, making it suitable for pooled functional genomics screens and population-level chromatin analyses. Users can interrogate the collective impact of H2BC12 ablation on nucleosome dynamics and global chromatin architecture in a cancer-relevant background.
HeLa cells are a cervical adenocarcinoma-derived line immortalized by human papillomavirus type 18 (HPV18) E6 and E7 oncoproteins, resulting in functional inactivation of the tumor suppressors p53 and retinoblastoma protein (Rb). This immortalized epithelial model exhibits aneuploidy, rapid proliferation, and dysregulated cell cycle control, characteristics that make it a standard host for studying chromatin biology and oncogenic transformation. The HPV18-driven background renders these cells particularly responsive to perturbations in histone supply and chromatin assembly, providing a sensitized system to observe phenotypes arising from H2BC12 deletion.
H2BC12 encodes histone H2B type 1-K, a core component of the nucleosome whose expression is tightly coupled to DNA replication via cell cycle-dependent transcriptional control. The H2BC12 promoter is activated by E2F transcription factors and the cyclin E/CDK2?CNPAT pathway at the G1/S boundary, ensuring adequate histone supply for nascent chromatin assembly. The encoded H2B protein interacts stoichiometrically with histones H2A, H3, and H4 to form the nucleosome octamer, and its deposition is facilitated by chaperones such as NAP1 and the FACT complex. H2BC12-containing nucleosomes are further modulated by ATP-dependent chromatin remodelers, influencing higher-order chromatin compaction, gene expression, and DNA repair. Disruption of H2BC12 therefore compromises nucleosome stability and perturbs the structural integrity of chromatin, with downstream effects on transcriptional regulation and genome maintenance.
In the HeLa context, H2BC12 knockout introduces a specific stress on the chromatin landscape that is superimposed on existing defects in cell cycle and DNA damage checkpoints. The absence of functional p53 and Rb pathways may lead to exacerbated genomic instability, altered replication fork progression, and adaptive shifts in global histone modification patterns. This model enables dissection of the specific contributions of replication-dependent histone H2B variants to chromatin organization and epigenetic signaling in a transformed cell environment. Researchers can explore how histone supply imbalances influence cancer cell fitness, gene expression heterogeneity, and sensitivity to DNA-damaging agents within a well-characterized oncogenic background.
This polyclonal knockout population is suited for a broad range of experimental approaches in chromatin biology, epigenetic regulation, and cancer research. Typical applications include western blotting to confirm loss of H2BC12 protein, RT-qPCR for transcript quantification, ChIP-seq to map genome-wide alterations in histone modifications, immunofluorescence to visualize nuclear organization, and RNA-seq to profile transcriptional consequences. Functional assays such as cell viability measurements and comet assays permit assessment of DNA damage accumulation. These cells also enable pooled CRISPR screens and drug response studies. For additional details or customization options, please contact Ascent Research.
