The BRD8 Knockout 786-O Polyclonal Cells product provides a heterogeneous pool of 786-O renal clear cell carcinoma cells with CRISPR/Cas9-mediated disruption of the BRD8 gene, creating a polyclonal loss-of-function model. This format reflects diverse editing outcomes across the population, suitable for functional analysis of BRD8-dependent pathways without clonal bias.
The 786-O cell line is a classical ccRCC model derived from a primary clear cell adenocarcinoma; it harbors a VHL mutation that inactivates pVHL, leading to constitutive HIF stabilization and oncogenic signaling. This genetic background makes it particularly suitable for analyzing the interplay between chromatin regulators and the VHL/HIF axis in renal cell carcinoma.
BRD8 encodes a bromodomain-containing scaffolding protein that is an integral subunit of the NuA4/TIP60 histone acetyltransferase (HAT) complex. This complex, which also includes EP400, TRRAP, and the catalytic subunit TIP60 (KAT5), catalyzes acetylation of histones H4 and H2A to modulate chromatin structure and transcriptional regulation. BRD8 functions downstream of ATM and ATR kinases, which are activated by DNA double-strand breaks; it facilitates recruitment of the NuA4 complex to damage sites, promoting histone acetylation and homologous recombination repair. BRD8 also interacts with p53, thyroid hormone receptor, and retinoic acid receptor, and it influences the transcriptional regulation of p21/CDKN1A and BAX, thereby linking chromatin remodeling to cell cycle control and apoptosis. Additionally, BRD8 associates with RUVBL1 and RUVBL2, further connecting it to chromatin remodeling and DNA repair.
In the context of VHL-deficient 786-O cells, BRD8 knockout is anticipated to impair NuA4/TIP60-dependent histone acetylation at DNA damage sites, compromising homologous recombination repair and potentially leading to genomic instability and sensitivity to DNA-damaging agents. Disruption of BRD8 may also alter the transactivation of p53 target genes such as p21 and BAX, thereby affecting cell cycle checkpoints and apoptotic responses. Consequently, this model provides a valuable system to dissect synthetic lethal interactions between chromatin acetylation and tumor suppressor pathways in ccRCC, and it may help uncover therapeutic vulnerabilities relevant to renal cell carcinoma.
This polyclonal knockout population supports a broad array of research applications. Direct detection of BRD8 protein levels via western blotting and assessment of histone H4 and H2A acetylation by ChIP-qPCR are feasible. DNA damage repair capacity can be measured using comet assays and ??H2AX immunofluorescence, while cell proliferation and apoptosis assays (MTT, colony formation) evaluate functional consequences. Co-immunoprecipitation experiments can verify interactions with complex members such as EP400, TRRAP, or TIP60, and RNA-sequencing can delineate transcriptomic changes. Additionally, HDAC inhibitor sensitivity testing may reveal epigenetic drug sensitivities in a BRD8-deficient background. These approaches make this model suitable for chromatin biology, DNA repair, functional genomics, and drug target validation studies. Researchers interested in obtaining this model are encouraged to contact Ascent Research for further technical details and ordering information.
