The BRD8 Knockout UM-UC-3 Polyclonal Cells product provides a heterogeneous population of UM-UC-3 human urothelial carcinoma cells with CRISPR/Cas9-mediated disruption of the BRD8 gene. As a polyclonal knockout pool, this reagent avoids clonal selection biases and captures the diversity of editing outcomes, offering a physiologically relevant loss-of-function model. CRISPR/Cas9-induced double-strand breaks lead to a mixed population of BRD8-null alleles, collectively abolishing protein expression. This model is suited for studying BRD8-dependent processes in a context that mirrors tumor heterogeneity.
The parental UM-UC-3 line originates from a primary human bladder transitional cell carcinoma and harbors a TP53 mutation, representing an invasive urothelial carcinoma model. These cells are extensively used to investigate bladder cancer biology, displaying characteristic invasive properties. The TP53-mutant genetic background provides a relevant setting to examine how additional disruption of chromatin regulators such as BRD8 modulates cancer cell behavior and treatment responses.
BRD8 is a bromodomain-containing scaffold protein integral to the NuA4/TIP60 histone acetyltransferase complex. This complex is recruited to sites of DNA damage by ATM and ATR kinases and interacts with p53, TRRAP, TIP60/KAT5, and EP400 to acetylate histone H4. BRD8-mediated histone acetylation facilitates p53-dependent transcriptional activation of target genes including p21/CDKN1A, PUMA, and BAX. Through this mechanism, BRD8 integrates DNA damage signals with chromatin remodeling to regulate cell cycle arrest and apoptosis. Consequently, BRD8 functions as a critical node linking the DNA damage response, histone modification, and p53 tumor suppressor pathways.
In the TP53-mutant UM-UC-3 context, BRD8 knockout can further impair residual p53-dependent transcription and DNA damage responses, allowing dissection of both p53-dependent and -independent NuA4/TIP60 functions. The polyclonal knockout population enables robust assessment of average gene disruption effects without clonal drift, making it suitable for studying alterations in downstream effectors such as p21 and PUMA. This model provides a platform to investigate how chromatin modifications influence invasive phenotypes and sensitivity to genotoxic agents in bladder cancer.
These polyclonal knockout cells support a range of assays: western blotting for p53 and acetylated histone H4; RT-qPCR for p21 and PUMA; ChIP-qPCR for histone acetylation at p53 target promoters; RNA-seq for transcriptome-wide analysis; flow cytometry for apoptosis and cell cycle profiling; migration and invasion assays; and drug sensitivity testing with chemotherapeutics such as cisplatin. This product is a versatile tool for research into p53 signaling, chromatin biology, and bladder cancer therapeutics. For further technical details, contact Ascent Research.
