Cat.No. : AAV00176Z
Titer: ≥1x10^12 GC/mL / ≥1x10^13 GC/mL Size: 30 ul/100 ul/500 ul/1 ml
Serotype: AAV Serotype 4 Storage: -80 ℃
| Cat. No. | AAV00176Z |
| Description | AAV serotype 4 particles contain GFP under CMV promoter. |
| Reporter | GFP |
| Serotype | AAV Serotype 4 |
| 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 AAV (adeno-associated virus) gene therapy is a promising approach to treat inborn errors of metabolism, but questions remain about its efficacy and durability. Studies have shown that tolerogenic ImmTOR nanoparticles coated with rapamycin prevent the formation of neutralizing anti-capsid antibodies, allowing vector re-administration. Here, researchers further demonstrate that ImmTOR mixed with AAV vector also enhances hepatic transgene expression at the initial dose of AAV vector, independent of its effects on adaptive immunity. ImmTOR enhances AAV trafficking to the liver, resulting in increased hepatic vector copy number and transgene mRNA expression. Enhanced transgene expression is achieved by an AAV receptor (AAVR)-independent mechanism that cannot be replicated in vivo with free rapamycin or empty nanoparticles. The multi-pronged mechanism of ImmTOR action makes it an attractive candidate to achieve more efficient transgene expression at the first dose while suppressing the adaptive response to AAV, allowing for repeated dosing.
Expression of AAVR in Huh-7 cells was knocked down by treatment with small interfering RNA (siRNA) specific for AAVR (Figure 1A). Huh-7 cells expressing approximately 20% of normal AAVR (Figure 1A) had a 50% reduction in AAVAnc80-luciferase expression (Figure 1B). However, mixing AAVAnc80-luciferase with ImmTOR restored transduction of Huh-7 cells treated with AAVR siRNA and increased luciferase expression by 35% compared to untreated cells (Figure 1B). These results suggest that ImmTOR may promote AAV transduction of hepatocytes in a manner independent of AAVR. To test this possibility, the researchers performed in vivo studies with AAV4, an AAV serotype that does not use AAVR. While liver transduction with AAV4-GFP alone was inefficient, with very low genome copy numbers observed in recipient mouse hepatocytes and no fluorescence detected above background levels, administration of AAV4-GFP mixed with ImmTOR resulted in higher viral genome copy numbers and detectable GFP fluorescence in hepatocytes of recipient animals (Figure 1C).
Figure 1. ImmTOR restores AAV8-Luc transgene expression after DsiRNA knockdown of AAVR in vitro and enhances AAV4 transduction in vivo. (Ilyinskii P O, et al., 2021)
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Beyond the excellent performance of the AAV4-GFP product itself, I have been thoroughly impressed by the company’s customer service. Their support team provided detailed guidance throughout the setup process and was quick to address any queries.
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