Cat.No. : AAV00178Z
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. | AAV00178Z |
| Description | AAV serotype 4 particles contain Cre recombinase under CMV promoter. |
| Serotype | AAV Serotype 4 |
| 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 using adeno-associated virus (AAV) vectors requires knowledge of their tropism in vivo. Here, researchers analyzed the tropism of ten naturally occurring AAV serotypes (AAV3B, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAVrh8, AAVrh10, and AAVrh74) after systemic delivery into male and female mice. Transgenes expressing ZsGreen and Cre recombinase were used to identify transduced cells in a cell-dependent manner based on fluorescence. Cre-driven tdTomato fluorescence activation provided excellent sensitivity for transduced cells. All serotypes, except AAV3B and AAV4, had high liver tropism. Fluorescence activation revealed transduction of unexpected tissues, including adrenal glands, testes, and ovaries. Rare transduced cells within tissues were also readily observed. Biodistribution of the AAV genome correlated with fluorescence, with the exception of immune tissues. The study found that AAV4 has pan-endothelial tropism and also targets pancreatic β cells.
Here, researchers generated AAV4 vectors expressing Cre recombinase. Intravenous injection of AAV4-Cre into Ai9 mice resulted in abundant tdTomato-positive signals in the lungs outside the airways (Figure 1A). Flow cytometry showed that the majority of tdTomato signals originated from endothelial cells based on CD31+/CD45- expression. Notably, >85% of lung endothelial cells were transduced by AAV4 in both male and female mice. In contrast, only about 20% of non-immune non-endothelial cells in the lung appeared to be transduced by AAV4 (Figure 1B). Repeating the experiment using an AAV4 vector expressing Cre demonstrated poor AAV4 transduction of the liver, showing only sparse TdTomato hepatocytes and possible signal in isolated liver sinusoidal endothelial cells. In stark contrast, impressive endothelial cell transduction was observed in the stomach, bladder, adrenal glands, pancreas, brain, small intestine, and thymus (Figure 1C). Furthermore, co-staining with insulin antibodies confirmed that AAV4 could effectively transduce β cells (Figure 1D).
Figure 1. AAV4 exhibits a unique tropism for endothelial cells and pancreatic beta cells. (Walkey C J, et al., 2024)
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AAV4-Cre's ability to provide clear, localized expression of Cre recombinase has greatly enhanced our research outcomes, avoiding off-target effects and ensuring reliable data. Highly recommended!
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