The BRD8 Knockout KYSE-30 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population offering a robust model for studying BRD8 loss-of-function. The polyclonal format provides a heterogeneous pool of gene-disrupted cells, avoiding clonal variation and enabling immediate large-scale functional assays without single-cell isolation.
KYSE-30 is a well-characterized human esophageal squamous cell carcinoma (ESCC) cell line derived from a well-differentiated primary tumor. ESCC is a deadly malignancy with limited therapeutic options; KYSE-30 retains hallmark genetic alterations and epithelial features, serving as a standard model for cancer biology and drug development in esophageal research.
BRD8 serves as a core subunit of the NuA4/TIP60 histone acetyltransferase complex, catalyzing acetylation of histones H2A and H4 as well as non-histone substrates including p53 and ATM. This acetylation relaxes chromatin and modulates protein function, driving transcriptional activation of DNA repair genes (GADD45, p21). BRD8??s bromodomains recognize acetylated lysine residues, orchestrating complex recruitment. The complex is regulated by upstream kinases ATM/ATR and transcription factor p53 in response to DNA damage. Critical interacting partners encompass TIP60 (KAT5), EP400, TRRAP, DMAP1, RUVBL1, RUVBL2, and ACTL6A. Disruption of BRD8 impairs acetylation dynamics, leading to defective DNA damage signaling and aberrant gene expression.
Esophageal squamous cell carcinoma exhibits pronounced genomic instability, often linked to compromised DNA repair pathways. BRD8??s function in the NuA4/TIP60 complex positions it at the nexus of chromatin biology and genome maintenance. This knockout model in KYSE-30 cells enables dissection of BRD8-dependent mechanisms that sustain tumor cell survival under genotoxic stress. It is particularly suited for synthetic lethality screens with DNA-damaging agents such as cisplatin, as well as for investigating epigenetic contributions to therapeutic resistance and p53 pathway interactions.
Applications encompass functional genomics, cancer epigenetics, and DNA damage response interrogation. Typical assays include western blot for acetyl-H2A/H4 and p53 acetylation, RT-qPCR for p21 and GADD45, and ChIP-qPCR for histone H4 acetylation. ??H2AX immunofluorescence monitors DNA breaks; flow cytometry profiles cell cycle and apoptosis. Clonogenic survival and cisplatin sensitivity assays evaluate drug responses, while RNA-seq and co-immunoprecipitation probe transcriptomic changes and complex integrity. For further technical information, please contact Ascent Research.
