Cat.No. : AD00138Z
Titer: ≥1x10^10 IFU/mL / ≥1x10^11 IFU/mL / ≥1x10^11 VP/mL / ≥1x10^12 VP/mL Size: 100 ul/500 ul/1 mL
Storage: -80℃ Shipping: Frozen on dry ice
| Cat. No. | AD00138Z |
| Target Gene | CEBPA |
| Product Type | Adenoviral particle |
| Insert | CEBPA |
| Titer | Varies lot by lot, for example, ≥1x10^10 IFU/mL, ≥1x10^11 IFU/mL, ≥1x10^11 VP/mL etc. |
| Size | Varies lot by lot, for example, 250 ul, 500 ul, 1 mL etc. |
| Storage | Store at -80℃. Avoid multiple freeze/thaw cycles. |
| Shipping | Frozen on dry ice |
| Creative Biogene ensures high-quality adenovirus particles by optimizing and standardizing production protocols and performing stringent quality control (QC). The specific QC experiments performed vary between adenovirus particle lots. | |
| Endotoxin | Endotoxins, primarily derived from Gram-negative bacteria, can trigger adverse immune responses. Endotoxin contamination is a significant concern in adenovirus production, especially for applications in animal studies and gene therapy. Creative Biogene utilizes rigorous endotoxin detection methods to monitor the endotoxin level in our produced adenovirus particles to ensure regulatory compliance. |
| Sterility | Creative Biogene ensures that adenovirus products are free of any bacterial, fungal and other microbial contamination. |
| Ad5 E1 Detection | All Creative Biogene adenoviruses are PCR tested to ensure that there are no detectable E1 sequences in the particles, which could be from revertants or external E1 contamination. |
| RCA Assays | Adenovirus products originating at Creative Biogene are guaranteed to have undetectable replication-competent adenovirus (RCA). This quality control measure is important because there is always the possibility of wild-type contamination due to revertants or environmental sources. |
| PFU Titering | All purified adenovirus preparations are tested for infectious titer. Creative Biogene's PFU test takes a few days longer but counts true plaques in HEK cells rather than estimating PFU titers via IHC staining or TCI50 of infected cells. |
A common denominator of both type 1 and type 2 diabetes is a reduction in functional β-cell mass. The transcription factor Nkx6.1 regulates β-cell development and is integral to normal β-cell function. Previous studies have shown that Nkx6.1 is dependent on c-Fos-mediated upregulation and nuclear hormone receptors Nr4a1 and Nr4a3 to increase β-cell insulin secretion, survival, and replication. Here, researchers demonstrate that Nkx6.1 overexpression leads to upregulation of the bZip transcription factor CEBPA and that CEBPA expression is independent of c-Fos regulation. In turn, CEBPA overexpression is sufficient to enhance proliferation of INS-1 832/13 β-cells and primary rat islets. CEBPA overexpression also increases the survival of β-cells treated with thapsigargin. The increased survival in response to ER stress corresponds with changes in the expression of various genes involved in the unfolded protein response, including a decrease in Ire1a expression. These data suggest that CEBPA is sufficient to enhance functional β-cell mass by increasing β-cell proliferation and modulating the unfolded protein response.
To investigate the effects of CEBPA expression in beta cells, the researchers used a CEBPA overexpressing adenovirus and demonstrated that it was sufficient to increase CEBPA mRNA (Figure 1A) and protein levels (Figure 1B, C). It has been previously demonstrated that Nkx6.1 induces c-Fos expression, which in turn induces Nr4a1, Nr4a3, and VGF expression. Induction of these genes is sufficient to enhance beta cell proliferation, insulin secretion, and survival. Here, the researchers sought to determine whether CEBPA overexpression would induce expression of c-Fos, VGF, Nr4a1, and Nr4a3. CEBPA overexpression increased c-Fos mRNA, but not c-Fos protein (Figure 1D-F). CEBPA overexpression did not alter VGF mRNA or protein levels (Figure 1G-I). CEBPA decreased Nr4a1 mRNA levels, but did not alter protein levels (Figure 1J-L). Finally, CEBPA induced the expression of Nr4a3 at both mRNA and protein levels ( Figure 1M-O ). These data suggest that CEBPA overexpression is sufficient to induce Nr4a3 expression but does not increase the protein levels of other Nkx6.1 target genes, c-Fos, VGF, or Nr4a1.
Figure 1. CEBPA overexpression increases protein level of Nr4a3. (Ellsworth P N, et al., 2024)
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Creative Biogene’s CEBPA adenoviral particles performed exceptionally well in our experiments. The purity and activity were as described, making them a trusted choice.
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