This product is a pool of CRISPR/Cas9-edited polyclonal BAZ1B knockout HEK293T cells, generated by targeted disruption of the BAZ1B gene in the human embryonic kidney cell line HEK293T. The polyclonal population contains a heterogeneous mixture of loss-of-function alleles, providing a robust model for studying BAZ1B-dependent processes without clonal artifacts.
HEK293T cells are an immortalized, adherent line derived from HEK293 cells, stably expressing the SV40 large T antigen and adenovirus E1A. They are widely used for transient transfection and recombinant protein production due to their high transfectability. Importantly, these cells exhibit low endogenous BAZ1B expression, making them an ideal host for knockout studies to minimize background effects.
BAZ1B is a core subunit of the WICH chromatin remodeling complex, where its bromodomain recognizes DNA damage-induced histone H3 acetylation at lysine 14 (H3K14ac). This interaction recruits the ATPase SMARCA5 to reposition nucleosomes, facilitating access for repair factors. BAZ1B functions downstream of ATM kinase-mediated signaling at DNA double-strand breaks and promotes downstream events including H2AX phosphorylation (??H2AX) and PCNA-dependent repair synthesis. It also interacts with SIRT1, RNF20, and histone H3, linking chromatin dynamics to transcription regulation and cell cycle checkpoint control. Disruption of BAZ1B impairs homologous recombination repair and alters transcriptional programs, contributing to genome instability.
In the HEK293T background, BAZ1B knockout allows clear dissection of its roles in DNA damage response and chromatin remodeling, unperturbed by high basal expression. This model is particularly valuable for investigating the molecular pathogenesis of Williams-Beuren syndrome, a neurodevelopmental disorder caused by haploinsufficiency of BAZ1B, as well as its emerging roles in cancer. The cell line’s ease of transfection enables rescue experiments with wild-type or mutant BAZ1B constructs to validate functional domains. Furthermore, co-knockout or overexpression of interacting partners like SMARCA5 or PCNA can elucidate pathway hierarchy.
Researchers can employ these cells in a variety of assays, including ChIP-qPCR to assess histone modifications at repair sites, immunofluorescence for ??H2AX foci quantification, comet assays to measure DNA damage, and Western blotting for phospho-ATM and BAZ1B levels. Functional studies may include RT-qPCR of BAZ1B target genes, cell viability assays following genotoxic stress, and luciferase-based transcription reporter assays. The polyclonal pool is well-suited for high-content screening of DNA damage modulators and epigenetic drug testing. For further details or to discuss your specific application, please contact Ascent Research.
