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Reporter AAV Particles optimize AAV-mediated gene transfer by using reporter proteins such as luciferase or fluorescent markers. These ready-to-use viral particles are ideal for visualizing and/or quantifying gene expression after AAV transduction. Adeno-associated virus (AAVs) containing reporter genes such as GFP, mCherry, RFP, lacZ and luciferase can be used as internal controls to optimize transduction and experimental conditions, track transgene expression over time or follow AAV behavior and transduction in vivo.
Reporter AAV Particles have a wide range of applications for in vivo infection in animals. If you are not familiar with using AAV as a gene delivery tool, here are some things you should consider before you start:
Ideally, the promoter should match the promoter of the virus expressing the GOI. If your goal is broad expression, promoters with broad activity, such as CAG (chicken beta actin promoter with CMV immediate early enhancer), CMV, EF1a, EFFS, UBC, PGK is a good choice. If you have a specific target cell type, you may want to try a different promoter, such as Camk2a for neuron-specific promoters. you can make your choice depending on your experimental conditions.
Creative Biogene offers easy-to-use and detectable reporter genes, including GFP, mCherry, RFP, lacZ and luciferase.
Creative Biogene offers a variety of AAV serotypes, as different AAV serotypes have different tissue tropism in vivo. The common serotypes and their tropism are shown in the table below. You can choose the best AAV serotype for in vivo studies based on your experimental needs.
| Tissue | Optimal Serotype |
|---|---|
| Heart | AAV1, AAV8, AAV9 |
| Central Nervous System (CNS) | AAV1, AAV2, AAV4, AAV5, AAV8, AAV9 |
| Kidney | AAV2 |
| Liver | AAV7, AAV8, AAV9 |
| Lung | AAV4, AAV5, AAV6, AAV9 |
| Pancreas | AAV8 |
| Photoreceptor Cells | AAV2, AAV5, AAV8 |
| Skeletal Muscle | AAV1, AAV6, AAV7, AAV8, AAV9 |
| (Retinal Pigment Epithelium (RPE) | AAV1, AAV2, AAV4, AAV5, AAV8 |
Depending on your experimental needs, there are several factors associated with in vivo AAV delivery that can greatly affect the degree of success of the infection. Regardless of your application, listed below are some of the factors you may want to consider before proceeding with delivery:
The number of viral particles you can deliver to a tissue will depend on the titer of your viral preparation and the maximum volume that can be delivered, which is very important. You can first search the literature for recommendations for specific tissues, but it is often necessary to determine the optimal dose empirically in vivo by trying a series of dilutions.
First, it is important to remember that terminally differentiated cells (i.e., post mitotic cells) will express AAV for a long time, but in dividing cells, expression will be lost. The age of the injected animal will determine which type of cells you can successfully infect.
Before starting the delivery process, you should also consider how to visualize your rAAV infection. Creative Biogene's reporter AAV particles incorporate reporter genes, such as GFP or mCherry, by incorporating them into the rAAV construct, and these will make it easy to directly observe where expression occurs through fluorescence measurements. Some of the popular reporter AAV particles are listed below, contact us to learn more about the products.
| Product Name | Promoter | Reporter | Serotype |
|---|---|---|---|
| CAG-GFP Adeno-associated virus (AAV Serotype 9) | CAG | GFP | AAV9 |
| CAG-GFP Adeno-associated virus (AAV Serotype 8) | CAG | GFP | AAV8 |
| LacZ Adeno-associated virus (AAV Serotype 9) | CMV | LacZ | AAV9 |
| GFP Adeno-associated virus (AAV Serotype 8) | CMV | GFP | AAV8 |
| LacZ Adeno-associated virus (AAV Serotype 6) | CMV | LacZ | AAV6 |
| GFP Adeno-associated virus (AAV Serotype 6) | CMV | GFP | AAV6 |
| LacZ Adeno-associated virus (AAV Serotype 5) | CMV | LacZ | AAV5 |
| LacZ Adeno-associated virus (AAV Serotype 2) | CMV | LacZ | AAV2 |
| Synapsin-GFP AAV (Serotype 1) | Syn | GFP | AAV1 |
| mRFP AAV (Serotype 2) | CMV | mRFP | AAV2 |
Sufficient time is required between injection and tissue processing to detect AAV-mediated gene expression. The time depends greatly on the type of capsid and the tissue you infect. Waiting about 2 weeks is a good starting point for many tissues. AAV-mediated gene expression has been reported to be very stable, persisting for several years in human clinical trials and in dogs.
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