The H2AC25 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed for targeted disruption of the H2AC25 gene in a well-established human embryonic kidney epithelial background. This product provides a heterogeneous pool of edited cells, each carrying loss-of-function mutations within the H2AC25 locus, enabling robust functional studies without the clonal selection biases inherent to single-cell-derived lines. The polyclonal format preserves population-level heterogeneity while maintaining consistent gene knockout across the majority of cells, making it ideal for experiments requiring representative biological replicates and high-throughput phenotypic screening.
HEK293T cells are derived from human embryonic kidney tissue and stably express the SV40 large T antigen, which promotes episomal replication of transfected plasmids and drives high-level protein expression. This cell line is widely utilized for viral production, transient protein overexpression, and gene editing applications due to its high transfection efficiency and rapid proliferation. The epithelial origin and transformed nature of HEK293T provide a permissive context for studying fundamental cellular processes, including chromatin biology and DNA repair, under conditions that support robust experimental manipulation and biochemical readouts.
H2AC25 encodes a canonical replication-dependent histone H2A variant that forms the structural core of nucleosomes by assembling into octamers with histones H2B, H3, and H4. The H2AC25 protein wraps approximately 147 base pairs of DNA and regulates chromatin condensation, DNA accessibility, and transcriptional activity. H2AC25 expression is transcriptionally regulated by E2F transcription factors and the NPAT coactivator, acting downstream of Cyclin E/CDK2 signaling during S-phase entry. The encoded histone interacts directly with histone chaperones NAP1 and the FACT complex to facilitate nucleosome assembly and disassembly, and is a substrate for ATP-dependent chromatin remodeling complexes such as SWI/SNF. Through these interactions, H2AC25 plays a central role in modulating nucleosome positioning, gene expression programs, and the cellular response to DNA damage, with disruption of its function leading to altered chromatin states and global transcriptomic changes.
Knockout of H2AC25 in HEK293T cells creates a powerful loss-of-function model for dissecting core histone biology in a fast-growing, epigenetically accessible system. Because HEK293T cells exhibit a hyperdiploid karyotype and active replication, they are especially suited to studying replication-coupled histone deposition and the consequences of histone imbalance on genomic stability. The polyclonal knockout population enables investigation of H2AC25-dependent mechanisms in transcriptional regulation, chromatin architecture, and DNA repair under conditions that mirror the heterogeneity found in human diseases such as cancer and genomic instability syndromes.
This knockout cell product is applicable to a broad range of molecular and cellular assays including chromatin accessibility profiling by ATAC-seq, genome-wide mapping of histone modifications by ChIP-seq, transcriptomic analysis via RNA-seq, and protein-level validation by Western blotting and RT-qPCR. Cell cycle distributions can be assessed by flow cytometry, while DNA damage responses can be evaluated using standard repair kinetics assays. These applications facilitate research into epigenetic deregulation in cancer, the role of core histones in genome maintenance, and the interplay between chromatin structure and transcriptional control. For technical specifications, batch validation data, and ordering information, please contact Ascent Research.
