Cat.No. : AAV00498Z
Titer: ≥1x10^12 GC/mL / ≥1x10^13 GC/mL Size: 30 ul/100 ul/500 ul/1 ml
Serotype: AAV Serotype 2 Storage: -80 ℃
| Cat. No. | AAV00498Z |
| Description | AAV serotype 2 particles express firefly luciferase (FLuc) reporter gene under the control of CAG promoter. |
| Reporter | FLuc |
| Serotype | AAV Serotype 2 |
| 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. |
Intervertebral disc degeneration is the leading cause of low back pain and is currently the leading cause of disability worldwide. Adeno-associated virus (AAV)-based viral vectors have emerged as powerful transgene delivery vehicles, but their respective tropism, transduction efficiency, and relative toxicity have not been systematically studied in the intervertebral disc in vivo. Here, researchers used in vivo bioluminescence imaging to systematically compare multiple AAV serotypes, injection volumes, titers, promoters, and luciferase reporters to determine which result in high transduction efficiency of murine nucleus pulposus (NP) cells in vivo. AAV6 using the CAG promoter to drive transgene expression was found to provide excellent transduction efficiency/kinetics and low in vivo toxicity when delivered into NPs in mouse tail discs at a titer of 1011 GC/mL. This study also demonstrated that in vivo transduction of NP cells was significantly enhanced using a small cell permeabilizing peptide. Taken together, these data will contribute to basic and translational research on the successful use of AAV-mediated gene delivery for IVDs.
Here, researchers sought to determine how the titer of delivered AAV particles affects transduction efficiency and transgene expression kinetics in vivo in the mouse tail IVD. Mice were injected locally (3 μL) with AAV6-CAG-fLuc particles at titers of 106, 109, or 1011 GC/mL in their Co6/7 discs. BLi was then performed at 3, 7, 14, 21, 28, 35, 42, and 56 days following injection of AAV6-fLuc to determine the transduction efficiency and kinetics for each titer in vivo. A titer of 106 GC/mL did not produce a BLi signal at any time point tested, while a titer of 1011 provided superior results compared to 109 (Figure 1). The mean peak intensity of BLi for AAV6 was 52 P/s/mm2, which occurred earlier at 21 days, compared to the mean peak intensity on AAV2, which occurred later at 6 weeks (Figure 1A). Representative BLi images for 109 and 1011 titers (42-day time point) are shown in Figure 1B and C, respectively. In summary, a titer of 1011 GC/mL was used in all subsequent experiments.
Figure 1. Viral titer affects transduction efficiency and kinetics. (Kim C H, et al., 2022)
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By using Creative Biogene's AAV2-CAG-FLuc, we were able to precisely visualize and track gene expression patterns over time, significantly advancing our research into gene function.
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