Cat.No. : AAV00196Z
Titer: ≥1x10^12 GC/mL / ≥1x10^13 GC/mL Size: 30 ul/100 ul/500 ul/1 ml
Serotype: AAV Serotype 5 Storage: -80 ℃
| Cat. No. | AAV00196Z |
| Description | AAV serotype 5 particles contain mCherry under GFAP promoter for specific expression in astrocytes/glial cells. |
| Reporter | mCherry |
| Serotype | AAV Serotype 5 |
| Application | 1. Determination of optimal MOI (multiplicity of infection), administration methods etc. 2. Detection of the infection efficiency of the AAV serotype against a specific cell type or tissue. 3. Using reporter genes to visualize the distribution and expression of AAV vectors in live animals, helping assess the biodistribution and persistence of gene delivery. |
| Titer | Varies lot by lot, typically ≥1x10^12 GC/mL |
| Size | Varies lot by lot, for example, 30 μL, 50 μL, 100 μL etc. |
| Storage | Store at -80℃. Avoid multiple freeze/thaw cycles. |
| Shipping | Frozen on dry ice |
| Creative Biogene ensures high-quality AAV particles by optimizing and standardizing production protocols and performing stringent quality control (QC). The specific QC experiments performed vary between AAV particle lots. | |
| Endotoxin | Endotoxins, primarily derived from Gram-negative bacteria, can trigger adverse immune responses. Endotoxin contamination is a significant concern in the production of AAV, especially for applications in animal studies and gene therapy. Effective endotoxin quality control is essential in the development and manufacturing of AAV particles. Creative Biogene utilizes rigorous endotoxin detection methods to monitor the endotoxin level in our produced AAV particles to ensure regulatory compliance. |
| Purity | AAV purity is critical for ensuring the safety and efficacy of AAV-based applications.AAV capsids are composed of three main protein components, known as viral proteins: VP1, VP2, and VP3. These proteins play a critical role in the structure and functionality of the AAV capsid. Monitoring the VP1, VP2, and VP3 content in AAV preparations is essential for quality control in AAV production. Our AAV particles are tested for showing three clear bands of VP1, VP2 VP3 by SDS-PAGE. |
| Sterility | The AAV virus samples are inoculated into the cell culture medium for about 5 days to detect bacterial and fungal growth. |
| Transducibility | Upon requirement, Creative Biogene can perform in vitro or in vivo transduction assays to evaluate the ability of AAV to deliver genetic material into target cells or tissues, and assess gene expression and functional activities. |
| Empty vs. Full Capsids | Based-on our proprietary AAV production and purification technology, Creative Biogene can always offer AAV particles with high ratio of full capsids. If required, we can also assess the ratio for a specifc lot of AAV particles by transmission electron microscopy (TEM) or other methods. |
Sleep disruption is a common problem in the elderly and is often accompanied by low-grade chronic central and peripheral inflammation. Here, researchers investigated whether chronic neuroinflammation in the preoptic and basal forebrain area (POA-BF), a key sleep-wake regulatory structure, contributes to this disruption. They developed a targeted viral vector designed to overexpress tumor necrosis factor-α (TNFα), specifically in astrocytes (AAV5-GFAP-TNFα-mCherry), and injected it into the POA of young mice to induce increased neuroinflammation within the POA-BF. Compared to controls (treated with AAV5-GFAP-mCherry), mice with astrocyte TNFα overproduction within the POA-BF showed signs of increased microglial activation, indicating an aggravated local inflammatory environment. These mice also showed aging-like changes in sleep-wake organization and physical function, including (a) impaired sleep-wake function, characterized by disrupted sleep and wake during the day and dark phases, respectively, and a reduced ability to compensate for sleep loss; (b) dysfunction of VLPO sleep-active neurons, as evidenced by a decrease in neurons expressing c-fos after suvorexant-induced sleep; and (c) impaired physical function, as evidenced by decreased grip strength. These findings suggest that inflammation-induced dysfunction of sleep and wake regulation mechanisms within the POA-BF may be a key factor in sleep-wake disorders during aging.
Injection site and spread of viral vectors were verified by expressing mCherry under the control of the GFAP promoter. Figure 1 shows the injection of the viral vector AAV5-GFAP-TNFα-mCherry or its control virus AAV5-GFAP-mCherry in VLPO and the potential results of this approach. In the control and TNFα groups, the injected vector spread to a wider area (approximately 1-2 mm, depending on the volume) and transfected parts of the VLPO, medial POA, dorsolateral POA, and BF, including the level of the diagonal band Limbs, septum, and cholinergic magnocellular areas. The sleep/wake profile of TNFα mice was significantly altered compared with controls. Therefore, the changes in sleep-wake patterns observed in this study are most likely caused by increased TNFα production, primarily within the POA and BF. Furthermore, viral vector spread was roughly equivalent in the control and TNFα groups. Therefore, the number of affected POA-BF astrocytes and neurons may be comparable in the TNFα and control groups.
Figure 1. Schematic of the procedure used for inducing TNFα expression/production in the astrocytes within the VLPO-BF. (Kostin A, et al., 2024)
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The AAV5-GFAP-mCherry vector has been a game-changer for our neurological research. It offers exceptional specificity in targeting astrocytes, and the expression of mCherry is both robust and consistent.
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