Cat.No. : AAV00047Z
Titer: ≥1x10^12 GC/mL / ≥1x10^13 GC/mL Size: 30 ul/100 ul/500 ul/1 ml
Serotype: AAV Serotype 8 Storage: -80 ℃
| Cat. No. | AAV00047Z |
| Description | Cre Adeno-associated virus(AAV Serotype 8)which express Cre recombinase under the control CMV promoter. This product used in the Cre-lox system as a genetic tool to generate site-specific recombination of DNA between loxP sites in cultured cells and animal experiments. |
| Serotype | AAV Serotype 8 |
| Target Gene | cre |
| 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. |
Adeno-associated virus (AAV)-mediated gene delivery systems have been shown to be an effective tool for genetic manipulation of the inner ear. For example, hair cells (HCs) and a variety of other cell types can be transduced by local injection of AAV into the inner ear. However, the application of AAV-mediated CRISPR/Cas9 gene editing approaches to the inner ear of adult mice has not been studied. Here, the researchers studied several AAV serotypes in both newborn and adult mice and found that AAV8 had the highest efficiency for transducing inner HCs. They then tested the ability of Cre-expressing AAV8 to activate Cas9 in floxed-Cas9 knock-in mice and observed significant Cas9 activation in the inner ears of both newborn and adult animals. Neither the AAV8 virus itself nor the surgical procedure used to deliver it caused any damage to the HCs or compromised normal hearing. These studies demonstrate that local injection of AAV8-Cre can induce Cas9 activation, allowing for safe and efficient gene editing in the inner ear, expanding the range of gene editing tools for regulating inner ear gene expression as part of efforts to rescue inherited hearing loss, initiate HC regeneration, or develop gene therapy technologies.
In adult mice, almost all the IHCs, as well as many SGNs, were transduced 4 weeks after injection of 1 μl of AAV8-Cre through the PCSS (Figure 1). Adult mice showed a higher efficiency of IHCs transduction in the apical, middle, and basal turns compared with injected neonatal Cas9-KI mice. No evidence of obvious HC death (<5%) was observed in any of the three turns after AAV injection. In the apical turn, AAV8-Cre infected all the IHCs and outer phalangeal cells and a few Hensen's cells (Figure 1b, b'). In the middle turn, all the IHCs and few outer phalangeal cells were tdTomato positive (Figure 1e, e'). At the basal turn, all the IHCs were infected. A few infected cells in the stria vascularis were also observed (Figure 1h, h'). SGN-infection efficiency was very high in all three turns, but no infected OHCs were observed (Figure 1).
Figure 1. Inner ear cell subtypes infected by AAV8-Cre injected into adult Cas9-KI mice. (Kang W, et al., 2020)
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