Cat.No. : AAV00288Z
Titer: ≥1x10^12 GC/mL / ≥1x10^13 GC/mL Size: 30 ul/100 ul/500 ul/1 ml
Serotype: AAV Serotype 9 Storage: -80 ℃
| Cat. No. | AAV00288Z |
| Description | AAV serotype 9 particles contain GFP under EF1a promoter. |
| Reporter | GFP |
| Serotype | AAV Serotype 9 |
| 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. |
AAV-PHP.eB depends on endothelial cells to highly transduce the central nervous system (CNS) and is widely used for intravenous gene therapy. Here, researchers tested the transduction profile of AAV-PHP.eB and developed intravenous NeuroD1 gene therapy to treat ischemic stroke in mice. The study found that the AAV-PHP.eB-GFP control virus crossed the BBB and effectively infected brain cells in the normal brain. However, after stroke, AAV-PHP.eB-GFP control virus was highly restricted in the blood vessels. Surprisingly, after switching to the therapeutic vector AAV-PHP.eB-NeuroD1-GFP, the viral vector successfully crossed the blood vessels and infected brain cells. Using Tie2-cre transgenic mice, researchers demonstrated that NeuroD1 regulates endothelial gene expression to regulate AAV-PHP.eB transduction. NeuroD1 follows the changes in endothelial cell signaling pathways, effectively protects the integrity of the blood-brain barrier, reduces neuroinflammation, inhibits neuronal apoptosis, and rescues motor dysfunction after ischemic stroke. In addition, overexpression of NeuroD1 in brain cells further promotes neuroregeneration.
To develop non-invasive gene therapy for ischemic stroke, the researchers used two commonly used BBB-penetrating AAV vectors (AAV-PHP.eB and AAV9) to deliver the desired gene. The ubiquitous promoter EF-1α was used to drive the expression of the reporter gene GFP in adult C57/B6 mouse brain cells (AAV-PHP.eB-EF1α-GFP or AAV9-EF1α-GFP), the virus was injected intracranially into the striatum, and GFP expression was examined by immunostaining 7 days after virus injection (dpi). The results showed that most of the GFP-transduced cells were NeuN+ neurons in the striatum (Figure 1), indicating that in the normal adult mouse brain, the EF-1α promoter mainly drives the expression of GFP in neuronal cells when injected directly into the brain.
Figure 1. AAV-PHP.eB and AAV9 mainly target neurons in normal adult mice via intracranial injection. (He X, et al., 2023)
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We have been using the AAV9-EF1α-GFP vector in our neuronal studies and the expression levels have been outstanding. The product arrived well-packaged with clear instructions, making the entire process smooth and efficient.
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