The H2AC11 Knockout HEK293T Polyclonal Cells comprise a CRISPR/Cas9-edited polyclonal knockout cell population generated from HEK293T cells, designed for the disruption of the H2AC11 gene. This pool of edited cells provides a loss-of-function model for studying H2A clustered histone 11, a core nucleosomal histone variant. The polyclonal population reflects a heterogeneous set of gene disruptions mediated by CRISPR/Cas9, enabling the investigation of H2AC11 function without the bottlenecks of single-cell cloning. This product serves as a versatile tool for probing the roles of H2A histones in chromatin structure, DNA packaging, and gene regulation.
The parental HEK293T line is a widely used human embryonic kidney epithelial cell derivative that stably expresses the SV40 large T antigen. This feature enhances episomal replication of transfected plasmids, making HEK293T cells highly transfectable and ideal for transient protein expression and lentiviral packaging. The cells maintain a robust capacity for post-translational modifications and chromatin assembly, providing a suitable background for interrogating histone functions. Their rapid growth and well-characterized proteome facilitate biochemical and genomic assays. The HEK293T background supports the study of both replication-dependent and -independent histone incorporation pathways, offering a relevant context for assessing H2AC11-dependent chromatin dynamics.
H2AC11 encodes a replication-dependent H2A histone variant that forms the nucleosome core particle together with H2B, H3, and H4 histones. It is regulated by canonical cell cycle transcription factors such as E2F and NPAT, with its expression tightly coupled to DNA replication. Once synthesized, H2AC11 is assembled into chromatin by histone chaperones including NAP1 and FACT, and its incorporation influences nucleosome stability and positioning. Chromatin remodelers of the SWI/SNF family interact with H2A-containing nucleosomes to modulate DNA accessibility. Knockout of H2AC11 disrupts these interactions, potentially altering genome-wide histone modification patterns and downstream gene expression programs. The mechanistic consequences include changes in chromatin compaction and replication fidelity, affecting transcriptional output and epigenetic memory.
In HEK293T cells, loss of H2AC11 can unmask compensatory mechanisms involving alternative H2A variants, providing insight into histone code plasticity. The knockout model allows researchers to dissect the specific contribution of H2AC11 to nucleosome architecture in a physiologically active but non-cancerous epithelial context. Because HEK293T cells retain functional chromatin regulatory machinery, the impact on pathways such as DNA replication and cell cycle progression can be directly examined. The polyclonal nature of the population reduces clonal selection artifacts, ensuring a broader representation of gene inactivation events. This model is particularly valuable for studying epigenetic mechanisms underlying diseases linked to chromatin defects, including developmental disorders and cancer.
Typical applications include chromatin immunoprecipitation followed by quantitative PCR (ChIP-qPCR) to map H2A variant distribution, transcriptomic profiling via RNA-seq to identify differentially expressed genes upon H2AC11 loss, and Western blotting to assess histone modification changes. Immunofluorescence microscopy enables visualization of chromatin structure alterations, while flow cytometry can monitor cell cycle perturbations. The cells are suited for functional rescue experiments with wild-type or mutant H2AC11 constructs, facilitating structure-function analyses of the histone fold domain. Researchers investigating nucleosome assembly, histone chaperone interactions, or chromatin remodeler targeting will find this polyclonal knockout population a robust resource. For further technical details or bulk orders, please contact Ascent Research.
