The H2AC14 Knockout HeLa Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from HeLa cells, with disruption of the H2AC14 gene encoding a core histone H2A. This heterogeneous pool contains diverse editing events at the target locus, resulting in loss of H2AC14 protein. The polyclonal format minimizes clonal selection bias and faithfully represents the spectrum of knockout phenotypes across the cell population, making it suitable for studies of histone depletion effects on chromatin.
HeLa cells are an immortalized, HPV18-positive epithelial cell line derived from a cervical adenocarcinoma, characterized by aneuploidy and active telomerase. They serve as a standard model for human epithelial cell biology, cancer mechanisms, and pharmacological studies. The extensive use of HeLa cells in chromatin research and the availability of established protocols make them an ideal host for generating H2AC14 knockout cells, facilitating investigation of nucleosome structure and function in a cancer-relevant background.
H2AC14 is a canonical core histone essential for nucleosome assembly and chromatin organization. It interacts with histones H2B, H3, H4, and chaperones including NAP1L1, the FACT complex (SUPT16H, SSRP1), and CAF-1 (CHAF1A, CHAF1B, RBBP4). H2AC14 expression is controlled by the NPAT/HiNF-P complex downstream of Cyclin E/CDK2 and E2F transcription factors, coupling histone production to DNA replication. Knockout of H2AC14 disrupts nucleosome integrity, alters chromatin architecture, and impairs interactions with modifiers such as EZH2 and HDACs, leading to altered gene expression and compromised DNA damage responses.
In HeLa cells, H2AC14 depletion models histone insufficiency resembling chromatin dysregulation in cancer. Loss of this core histone reduces nucleosome occupancy, promoting genomic instability and aberrant transcription. This system is particularly useful for studying oncogenic mechanisms driven by altered histone stoichiometry, as seen in cervical and other malignancies. The polyclonal population captures a range of knockout effects, enabling robust analysis of cell cycle defects, replication stress, and sensitivity to genotoxic agents without clonal adaptation artifacts.
Applications include analyzing histone dosage effects on chromatin dynamics, nucleosome assembly, and genome stability using techniques such as MNase-seq, chromatin fractionation, and RNA-seq. The knockout cells are also suited for cell cycle profiling by flow cytometry, western blotting of histone complexes, co-immunoprecipitation of interaction partners, Comet assays for DNA damage, and proliferation assays to screen synthetic lethal interactions or epigenetic drug responses. For further information, contact Ascent Research.
