Cat.No. : AAV00182Z
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. | AAV00182Z |
| Description | Self-complementary AAV serotype 5 particles contain GFP under the control of CMV promoter. |
| Reporter | GFP |
| Serotype | AAV Serotype 5 |
| 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. |
CD40 ligand (CD40L) gene therapy has multiple antitumor activities and provides a potentially useful option for lung cancer. However, membrane-bound CD40L may be proteolytically cleaved to form soluble CD40L (sCD40L), resulting in adverse effects. Previous studies have demonstrated that recombinant self-complementing adeno-associated virus 5 (scAAV5) can efficiently deliver genes into lung cancer cells. Here, researchers generated an scAAV5 expressing a non-cleavable human CD40L mutant (scAAV5-CD40L-M) and evaluated its direct antitumor effects in lung cancer. Compared with scAAV5-CD40L transduction, scAAV5-CD40L-M transduction resulted in effective expression of CD40L on the cell surface with low levels of cleaved sCD40L, which significantly reduced the percentage of viable cells and promoted caspase-3-dependent apoptosis of CD40-positive lung carcinoma A549 cells. Furthermore, scAAV5-CD40L-M treatment exerted significant antitumor effects on CD40-positive A549 xenografts by inducing apoptosis with few side effects. Therefore, gene therapy using scAAV5 vectors expressing non-cleavable human CD40L mutants may have direct antitumor effects in CD40-positive lung cancer.
To focus on the direct tumor growth inhibitory properties of AAV5-CD40L-M, in vivo analyzes were performed in athymic nude mice via lung cancer xenografts. Both scAAV5-CD40L and scAAV5-CD40L-M effectively reduced tumor growth rate (Figure 1A). The average tumor weights for the scAAV5-GFP, scAAV5-CD40L, and scAAV5-CD40L-M groups were 0.43±0.03, 0.31±0.02, and 0.29±0.03 g, respectively (Figure 1B). CD40L expression was detected in scAAV5-CD40L- and scAAV5-CD40L-M-injected tumors but not in scAAV5-GFP-injected tumors (Figure 1C). The effect of scAAV5-CD40L-M treatment on apoptosis was assessed using an in situ TUNEL assay. The most significant levels of apoptosis were observed in tumors from mice treated with scAAV5-CD40L or scAAV5-CD40L-M compared with scAAV5-GFP mice (Figure 1D).
Figure 1. scAAV5-CD40L-M inhibits tumor growth in vivo. (Xu W, et al., 2015)
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The transduction efficiency is notably high, and the GFP signal remains strong over extended periods, facilitating long-term experiments without any signal degradation.
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