The H2AC4 Knockout HEK293T Polyclonal Cells provide a CRISPR/Cas9-mediated gene-disrupted population of human embryonic kidney cells, engineered to lack the replication-independent histone variant H2AC4 through polyclonal editing. This product consists of a heterogeneous pool of cells with targeted disruption of the H2AC4 gene, offering a physiologically relevant loss-of-function model without single-cell clonal selection. The polyclonal format preserves population-level diversity and is well suited for bulk biochemical and functional assays where clonal uniformity is not required, enabling robust investigation of H2AC4’s role in chromatin dynamics and gene regulation.
The host cell line, HEK293T, is a widely used derivative of the HEK293 parental line that stably expresses the SV40 large T antigen. This modification promotes high-copy episomal replication of plasmids containing the SV40 origin of replication, making HEK293T an exceptional system for recombinant protein overexpression and lentiviral or retroviral vector production. Its human embryonic kidney epithelial origin provides a relevant cellular environment for studying histone biology and epigenetic mechanisms, as these cells retain the core chromatin machinery and signaling networks essential for nucleosome assembly and DNA damage responses.
H2AC4 encodes a histone H2A variant that is synthesized and incorporated into nucleosomes independently of DNA replication, distinguishing it from canonical S-phase-dependent histones. This variant integrates into chromatin through the action of dedicated histone chaperones such as NAP1L1 and the FACT complex, and it interacts intimately with core histone partners H2B and linker histone H1. Its deposition is modulated upstream by DNA damage response kinases ATM and ATR, linking H2AC4 to surveillance pathways that govern genomic stability. Downstream, H2AC4 influences chromatin compaction and the accessibility of transcriptional regulators, and it participates in the recruitment of DNA repair proteins to sites of damage, bridging epigenetic marks with the cellular response to genotoxic stress.
In the HEK293T background, disruption of H2AC4 is expected to perturb nucleosome stability and the dynamic exchange of histone variants, leading to altered chromatin architecture. This may manifest as shifts in gene expression profiles due to changes in the local recruitment of chromatin remodelers such as the SWI/SNF complex and polycomb repressive components, which are key representative factors in the pathways associated with H2AC4. The knockout model thus serves as a powerful tool to dissect how replication-independent histones contribute to transcriptional regulation and epigenetic maintenance in a rapidly dividing, transformation-competent cell system.
Researchers can employ this polyclonal knockout population for a range of advanced applications, including chromatin immunoprecipitation (ChIP-qPCR or ChIP-seq) to map genome-wide histone variant occupancy and histone modification landscapes, western blotting to evaluate changes in canonical and variant histone levels, and nucleosome assembly assays to examine chromatin dynamics in vitro. The model also supports RT-qPCR-based gene expression profiling and DNA damage response assays??such as monitoring ATM/ATR-dependent phosphorylation events or recruitment kinetics of repair factors??to elucidate H2AC4’s role in genome maintenance pathways. For further technical details or to discuss custom projects, please contact Ascent Research.
