The H2BC18 Knockout HEK293T Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the H2BC18 gene in human HEK293T cells. This polyclonal pool provides a loss-of-function model for studying the replication-dependent histone H2B protein without isolating individual clones. The heterogeneous collection of edited alleles enables assessment of global gene disruption effects across the population, offering a versatile tool for histone biology research in a widely used mammalian cell background. The knockout model is particularly suited for investigating how histone supply influences chromatin architecture and gene regulation.
HEK293T cells, the host line, are human embryonic kidney epithelial cells transformed with adenovirus 5 DNA and stably expressing the SV40 large T antigen. Renowned for high transfection efficiency and robust protein expression, HEK293T is a standard platform for viral production, recombinant protein synthesis, and fundamental cell biology studies. Their rapid proliferation and well-characterized nature provide a reliable context for examining chromatin dynamics and cell cycle processes, making them an ideal background for dissecting replication-dependent histone functions.
H2BC18 encodes a core component of the nucleosome, histone H2B, which is essential for packaging DNA into chromatin. This replication-dependent histone is synthesized during S-phase and incorporated into newly replicated DNA through interactions with histone H2A, H3, H4, and linker histone H1, as well as histone chaperones such as NASP and ASF1. Its expression is tightly regulated by the cell cycle machinery, including E2F transcription factors, NPAT coactivator, and Cyclin E/CDK2 signaling. Disruption of H2BC18 perturbs H2B supply, potentially altering nucleosome assembly and chromatin architecture, though compensatory histone variants may partially rescue function.
In the HEK293T background, H2BC18 knockout provides a valuable model to dissect the functional consequences of replication-dependent histone depletion. The polyclonal population may exhibit altered chromatin dynamics, impacting gene regulation programs and cell cycle progression, thereby revealing mechanisms of epigenetic control. This system allows investigation of interplay between histone dosage and epigenetic states and delineation of compensatory pathways involving other H2B isoforms, with relevance to how histone gene mutations contribute to oncogenic epigenetic dysregulation.
Researchers can employ these cells in applications such as western blotting to assess histone H2B and other histone levels, chromatin fractionation to monitor nucleosome assembly, RNA-seq for transcriptional profiling, immunofluorescence for chromatin organization, cell cycle analysis by flow cytometry, and apoptosis assays. The model is well-suited for studies in cancer epigenetics, histone variant compensation, and the impact of histone supply on gene expression. For additional information or technical support, please contact Ascent Research.
