The H2AC4 Knockout HeLa Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal cell population derived from HeLa cells, engineered for targeted disruption of the H2AC4 gene. This knockout model provides a loss-of-function system to investigate the roles of the H2A histone variant in chromatin biology. The polyclonal nature of the edited population reflects an ensemble of gene-disrupted alleles without single-cell clonal selection, enabling studies that capture a range of knockout effects.
HeLa cells are an immortalized human epithelial cell line originally isolated from a cervical adenocarcinoma. They harbor integrated human papillomavirus type 18 (HPV18) sequences and express viral oncoproteins, which contribute to their transformed phenotype and widespread use as a model in cancer biology. HeLa cells exhibit robust proliferation and are amenable to a wide range of genetic manipulation and phenotypic assays, making them a workhorse for studying cell cycle regulation, epigenetics, and drug response.
H2AC4 encodes a canonical core histone H2A protein that, together with histones H2B, H3, and H4, forms the nucleosome??the fundamental unit of chromatin. Histone gene expression is tightly coupled to cell cycle progression, regulated by the NPAT-HiNF-P complex and E2F transcription factors, and influenced by p53. H2AC4 interacts directly with H2B, H3, and H4 during nucleosome assembly, a process facilitated by histone chaperones such as NAP1 and ASF1. Mature nucleosomes serve as substrates for chromatin remodeling complexes, including the SWI/SNF complex, to modulate DNA accessibility. Disruption of H2AC4 can perturb nucleosome stoichiometry, potentially altering chromatin structure and transcription. This disruption may affect the dynamics of histone chaperone-mediated deposition and the activity of ATP-dependent remodelers.
In the HeLa cell context, H2AC4 knockout provides a powerful tool to dissect how core histone imbalances influence cancer cell phenotypes. HeLa cells, with their HPV-driven oncogenic program and aberrant cell cycle control, are particularly relevant for examining how chromatin disruption intersects with oncogenesis. Alterations in histone expression are observed in various cancers, and loss of H2AC4 may affect genome stability, gene expression programs, and responses to therapeutic agents. Thus, this model enables the study of histone-dependent mechanisms in a cervical cancer background.
These polyclonal knockout cells are suitable for a broad array of applications in epigenetics and cancer research. Researchers can employ chromatin immunoprecipitation sequencing (ChIP-seq) to map genome-wide changes in histone occupancy and modifications, or RNA-seq to assess transcriptional consequences. Cell cycle analysis by flow cytometry, proliferation assays, and apoptosis assays can reveal functional impacts on growth and survival. Immunofluorescence and western blotting facilitate validation of histone expression and nucleosome assembly defects. These cells are also valuable for drug response studies assessing the role of chromatin integrity in chemosensitivity. For additional technical details or ordering information, please contact Ascent Research.
