The H2AC14 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the human H2AC14 gene. H2AC14 encodes a replication-dependent core histone H2A variant that is integral to nucleosome formation and chromatin architecture. This polyclonal pool, generated in the HEK293T host cell background, provides a heterogeneous loss-of-function model for investigating the functional impact of H2AC14 loss. The polyclonal format averts clonal artifacts, facilitating the study of allele-level chromatin dynamics. These cells are well suited for applications in epigenetic biology, cancer modeling, and compound screening.
The HEK293T cell line is a human embryonic kidney epithelial line transformed with sheared adenovirus type 5 DNA and stably expressing the SV40 large T antigen. It is widely utilized for transient protein expression, viral packaging, and recombinant protein production due to its high transfection efficiency and rapid growth. Large T antigen expression enables episomal replication of plasmids containing the SV40 origin, making the line ideal for lentiviral and retroviral vector production. In chromatin studies, HEK293T cells exhibit active replication-coupled histone deposition, providing a relevant context for examining H2AC14 function.
H2AC14 is a replication-dependent core histone H2A variant that, with histones H2B, H3, and H4, forms nucleosomes. Its transcription is driven by E2F factors and the Cyclin E/CDK2 kinase complex via the NPAT coactivator at histone locus bodies. H2AC14 directly interacts with the FACT complex and NAP1L1 chaperone during nucleosome assembly and exchange, contributing to nucleosome stability and chromatin compaction. This, in turn, influences global gene expression patterns. Additionally, H2AC14 participates in DNA repair by recruiting repair machinery; its loss may compromise nucleosome stability and chromatin accessibility, thereby altering transcriptional and genomic maintenance programs.
Loss of H2AC14 in HEK293T cells provides a powerful model to probe histone variant functions in a transformed human epithelial line. The high proliferation rate and active DNA replication of HEK293T cells render them sensitive to alterations in histone supply and nucleosome assembly. The polyclonal knockout pool mirrors heterogeneous genetic backgrounds, allowing assessment of dosage effects and paralog compensation. This system is particularly relevant to cancer epigenetics, as histone variant deregulation is common in malignancies. Moreover, the presence of SV40 large T antigen, which interacts with host chromatin modifiers, adds a layer of complexity for studying viral oncoprotein effects on histone dynamics.
Applications include Western blotting to confirm histone expression changes, ChIP-qPCR for histone modifications, and immunofluorescence for chromatin structure analysis. Functional assays encompass cell cycle analysis by flow cytometry, DNA damage assays to measure repair capacity, and nucleosome positioning assays. The polyclonal knockout model also supports high-throughput screening for histone-modifying enzyme inhibitors or synthetic lethal partners. Specific studies may probe the role of H2AC14 in E2F-dependent transcription, FACT-mediated chromatin remodeling, or DNA repair pathway choice. For further information or custom project discussions, please reach out to Ascent Research.
