BRD3 Knockout Cell Line-293T
Cat.No. : CSC-RT0456
Host Cell: 293T Target Gene: BRD3
Size: 1x10^6 cells/vial, 1mL Validation: Sequencing
Inquire for Price
Cell Line Information
Cell Culture Information
Safety and Packaging
| Cat. No. | CSC-RT0456 |
| Cell Line Information | 293T-BRD3 (-/-) is a stable cell line with a homozygous knockout of human BRD3 |
| Target Gene | BRD3 |
| Host Cell | 293T |
| Species | Homo sapiens (Human) |
| Revival | Rapidly thaw cells in a 37°C water bath. Transfer contents into a tube containing pre-warmed media. Centrifuge cells and seed into a 25 cm2 flask containing pre-warmed media. |
| Mycoplasma | Negative |
| Format | One frozen vial containing millions of cells |
| Storage | Liquid nitrogen |
| Safety Considerations |
The following safety precautions should be observed. 1. Use pipette aids to prevent ingestion and keep aerosols down to a minimum. 2. No eating, drinking or smoking while handling the stable line. 3. Wash hands after handling the stable line and before leaving the lab. 4. Decontaminate work surface with disinfectant or 70% ethanol before and after working with stable cells. 5. All waste should be considered hazardous. 6. Dispose of all liquid waste after each experiment and treat with bleach. |
| Ship | Dry ice |
Inquiry
Background
Case Study
Applications
BRD3 (bromodomain-containing 3) is a protein-coding gene that belongs to the bromodomain and extra-terminal (BET) protein family, which also includes BRD2, BRD4, and BRDT. These proteins are characterized by the presence of two bromodomains and an extra-terminal (ET) domain, which enables them to recognize and bind to acetylated lysine residues on histone tails, thereby affecting chromatin structure and gene expression. BRD3 has been identified as a key player in the regulation of gene transcription. By binding to acetylated histones, BRD3 can recruit the transcriptional machinery to specific genomic regions, thereby promoting the transcription of certain genes. This regulatory function makes BRD3 an essential component of cellular processes such as cell signaling, growth, and differentiation.
Multiple studies have implicated BRD3 in various human diseases, most notably cancer. For example, BRD3 was found to form a fusion protein with the NUT gene (NUTM1), resulting in a malignant cancer called NUT midline carcinoma (NMC). This particular type of cancer is characterized by rapid and uncontrolled growth of cancer cells, often affecting midline structures such as the head, neck, and mediastinum. BRD3-NUT fusion proteins are thought to drive carcinogenesis by aberrantly maintaining chromatin in a hyperacetylated state, which in turn leads to uncontrolled gene expression and cell proliferation.
As members of the bromodomain and extra-terminal motif protein families, bromodomain-containing proteins regulate a variety of biological processes, including protein scaffolding, mitosis, cell cycle progression, and transcriptional regulation. The roles of these bromodomain proteins (Brds) in the innate immune response have been reported, but the role of Brd3 remains unclear. Researchers found that viral infection significantly downregulated Brd3 expression in macrophages, and Brd3 knockout inhibited virus-induced IFN-β production. Brd3 interacts with IRF3 and p300, increases p300-mediated IRF3 acetylation, and enhances the binding of IRF3 to p300 upon viral infection. Importantly, Brd3 promotes the recruitment of the IRF3/p300 complex to the Ifnb1 promoter and increases the acetylation of histone 3/histone 4 within the Ifnb1 promoter, thereby enhancing type I interferon production. Thus, these studies suggest that Brd3 may act as a coactivator for IRF3/p300 transcriptional activation of Ifnb1 and provide new insights into epigenetic mechanisms for efficient activation of innate immune responses.
Here, the researchers performed ChIP analysis to evaluate the effect of Brd3 on the recruitment of IRF3 and p300 to the Ifnb1 promoter. As shown in Figure 1a, virus-induced IRF3 and p300 binding to the Ifnb1 gene promoter was significantly reduced in Brd3 knockout (Brd3-ko) cells, indicating that Brd3 enhanced the virus-triggered recruitment of IRF3 and p300 to the Ifnb1 promoter. When Brd3 protein was knocked out, acetylated histone 3 and histone 4 within the Ifnb1 promoter were significantly reduced compared with control cells after virus infection (Figure 1b). However, the acetylation of histone 3 and histone 4 within the Tnfa promoter was not different between Brd3 knockout cells and control cells. Taken together, these results indicate that Brd3 promotes the transcriptional activity of IRF3/p300 by enhancing virus-triggered IRF3/p300 recruitment to the Ifnb1 promoter and assisting the acetylation of histone 3 and histone 4, thereby promoting the production of type I interferons.
Figure 1. Brd3 recruits IRF3/p300 complex to the promoter of Ifnb1 and facilitates the transcription of Ifnb1. (Ren, Wenhui, et al. 2017)
Creating a BRD3 knockout (KO) in the 293T cell line provides a powerful tool for studying the biological functions of BRD3 and its involvement in various cellular processes. Here are a few key applications of the BRD3 KO 293T cell line:
Gene Regulation Studies: BRD3 interacts with acetylated lysines on histone tails to influence gene expression patterns. The BRD3 KO 293T cell line can be used to study changes in gene expression by comparing the transcriptomes of wild-type and BRD3-deficient cells.
Cancer Research: The BET family of proteins, including BRD3, has been implicated in various cancers. The BRD3 KO 293T cell line can be used to understand the role of BRD3 in cancer cell proliferation, apoptosis, and metastasis.
Drug Screening and Development: By comparing the responses of wild-type and BRD3-deficient cells to various compounds, researchers can identify specific inhibitors of BRD3 and evaluate their therapeutic potential. This cell line can also be used to determine the specificity and efficacy of BET inhibitors and to study off-target effects of drugs targeting the BET family.
Epigenetic Regulation: As a member of the BET family, BRD3 plays an important role in epigenetic regulation of gene expression. Using the BRD3 KO 293T cell line, researchers can study the effects of BRD3 loss on the epigenetic landscape of cells.
Stem Cell Research: BRD3 has been implicated in the maintenance of stem cell pluripotency and differentiation. The BRD3 KO 293T cell line can be used to study the role of BRD3 in stem cell biology, including self-renewal and differentiation processes.
For research use only. Not intended for any clinical use.
