Cat.No. : AAV00507Z
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. | AAV00507Z |
| Description | AAV serotype 5 particles express firefly luciferase (FLuc) reporter gene under the control of CMV promoter. |
| Reporter | FLuc |
| Serotype | AAV Serotype 5 |
| Target Gene | FLuc |
| 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. |
Gene therapy has the potential to treat rheumatic diseases; however, macrophages in joints may impede adeno-associated viral vector-mediated gene delivery. Here researchers demonstrate that in arthritic, but also in healthy, mice administration of agents that influence macrophage activity/number and/or addition of empty decoy capsids substantially improve the efficacy of recombinant adeno-associated viral vector 5 transgene expression in the joint. Pretreatment with triamcinolone acetonide or clodronate liposomes improved luciferase expression within 4 weeks. Similar results were seen when empty bait capsids were added to capsid-containing whole genomes at a 5:1 ratio. In a study that evaluated the duration of expression and examined a combination of the two approaches, researchers observed a synergistic enhancement of gene expression that lasted for at least 12 weeks. Enhanced gene expression was independent of the route of administration of triamcinolone acetonide (intra-articular or intramuscular). In healthy mice, the results showed that the combination significantly improved transgene expression independent of serotype.
All previous experiments were performed in a CIA model in which animals develop severe inflammation in their joints when the vector is administered. Since even healthy synovium contains a significant proportion of macrophages, the researchers next decided to investigate whether empty decoy capsids could enhance gene expression in healthy joints. When empty capsids were added to genome containing particles at 2 different ratios (5:1, 20:1), there was a dose-dependent increase in gene expression (Figure 1). In animals injected with a 5:1 ratio of empty to full capsids, there was an average increase of 4.8-fold, while when a 20:1 ratio of empty to full capsids was used, there was a 20-fold increase.
Figure 1. Addition of empty capsids in 2 different ratios (5:1 and 20:1 empty to full) improves intra-articular rAAV5-luciferase expression in healthy mice. Healthy mice (n=7 mice per group, one joint injected per animal) were injected i.a. with 1.65e10 vg/joint rAAV5-CMV-FLUC. In 2 groups empty capsids were added in different ratios. Luciferase expression was measured weekly until mice were sacrificed after 4 weeks. (Aalbers C J, et al., 2017)
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