Cat.No. : AAV00254Z
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. | AAV00254Z |
| Description | AAV serotype 9 particles contain mCherry under CMV promoter. |
| Reporter | mCherry |
| 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. |
Systemic AAV9 delivery offers the potential for widespread and efficient retinal gene delivery and may therefore be a useful approach for treating diseases that are not amenable to intraocular injections, syndromes that affect multiple organs, or diseases that require early intervention. Expression produced by intravenous AAV9 is more effective in neonates than in adults, and here researchers describe the effects of age on AAV9 retinal transduction in the mouse retina. They found that the pattern of expression in neonatal mice correlated with retinal vascular development and that the areas of retinal transduction, as well as the cell types infected, varied depending on age at injection. Furthermore, sequential injections of AAV9 vectors carrying two different transgenes infected adjacent areas of the retina, providing greater coverage. Finally, these studies show that the spatiotemporal expression pattern of endogenous retinal Mfsd2a, a protein associated with maturation of a functional blood-brain barrier, is consistent with the inhibition of retinal transduction by intravenous AAV9, suggesting that AAV9 crosses the blood-brain barrier by endocytosis.
Systemic delivery of AAV to the mouse retina holds great promise for proof-of-concept gene therapy and basic studies of retinal development. However, results showed that a single injection transduced only a portion of the retina (determined by the age at the time of injection), limiting the effectiveness of this approach. Therefore, the researchers tested whether serial injections at two time points could be used to target a larger portion of the retina (Figure 1). Newborn mice (n = 5) were injected with AAV9-mCherry at P1 and then again with AAV9-GFP at P5. Using this approach, up to 2/3 of the retinal area could be routinely transduced. High-resolution images of the retina (Figure 1b, c) showed that P1 and P5 injections transduced non-overlapping populations of cells. Comparison of retinas injected at P1 alone, at P5 alone, or at both P1 and P5 showed that immune responses (including the production of neutralizing antibodies against the vector capsid) did not appear to interfere with expression of the second vector, likely due to the short time interval between injections (Figure 1d).
Figure 1. Sequential injections of AAV9-mCherry and AAV9-GFP cover an increased area of the retina. (Byrne L C, et al., 2015)
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Using AAV9-mCherry has expanded my research capabilities significantly. Its reliability and efficiency in gene delivery have allowed me to explore new avenues in gene expression studies that were previously limited by other methods.
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