The H2BC12 Knockout HEK293T Polyclonal Cells comprise a CRISPR/Cas9-edited polyclonal population of HEK293T cells with targeted disruption of the H2BC12 gene, which encodes a replication-dependent histone H2B. This heterogeneous knockout model avoids clonal selection, preserving natural cellular variability and providing a robust system for investigating histone function. The product is intended for advanced research in chromatin biology and epigenetic regulation.
HEK293T is a human embryonic kidney cell line immortalized with adenovirus 5 DNA and engineered to express SV40 large T antigen. This enables episomal plasmid replication and confers exceptionally high transfection efficiency, making it a standard host for molecular and cellular studies. The adherent epithelial cells are well-characterized and widely used for gene editing, protein expression, and functional assays.
H2BC12 is a core histone H2B synthesized during S phase under the control of E2F transcription factors and Cyclin E/CDK2, with mRNA processing regulated by SLBP and HINFP. The protein integrates into nucleosomes alongside H2A, H3, and H4, and its deposition is mediated by histone chaperones NAP1, FACT, and CAF-1. In chromatin, H2BC12 interfaces with chromatin remodelers like SWI/SNF and is dynamically modified by ubiquitin ligases RNF20/RNF40 and deubiquitinase USP22. This network governs chromatin compaction and accessibility for transcription, replication, and DNA repair.
In HEK293T cells, loss of H2BC12 can perturb nucleosome assembly and chromatin structure, providing a platform to dissect the contribution of H2B histones to cell cycle progression and gene regulation. The polyclonal nature allows observation of a range of knockout severities, and the high transfectability supports rescue with wild-type or mutant H2BC12 to validate phenotypic effects.
Key applications include Western blotting for histone modifications, immunofluorescence for chromatin architecture, and flow cytometry for cell cycle distribution. ChIP-qPCR can monitor nucleosome occupancy, while RNA-seq profiles transcriptomic shifts. This system is particularly valuable for cancer epigenetics research focusing on oncohistone mutations. For ordering and technical support, please contact Ascent Research.
