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Cre Adeno-associated virus(AAV Serotype 2)

Cre Adeno-associated virus(AAV Serotype 2)

Cat.No. :  AAV00045Z

Titer: ≥1x10^12 GC/mL / ≥1x10^13 GC/mL Size: 30 ul/100 ul/500 ul/1 ml

Serotype:  AAV Serotype 2 Storage:  -80 ℃

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AAV Particle Information

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Cat. No. AAV00045Z
Description Cre Adeno-associated virus(AAV Serotype 2)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 2
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.
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Adeno-associated virus (AAV), belonging to the parvovirus family, is a non-enveloped single-stranded linear DNA virus that requires a helper virus (usually adenovirus) to participate in replication. The genome of AAV is about 4800bp, with two ITR sequences and two open reading frames. AAV has many common serotypes and more than 100 virus variants. Among them, AAV2 is the most widely used adeno-associated virus and has good effects on most common cells. The natural tropism of AAV2 is skeletal muscle, neurons, and certain blood vessels and liver cells, which has stimulated people's interest in its use to treat a variety of diseases that affect these tissues. Gene therapy trials involving AAV2 have shown good prospects in treating diseases such as Leber's congenital amaurosis, hemophilia, and certain neurodegenerative diseases. Cre adeno-associated virus (AAV) serotype 2 is a widely used tool in molecular biology and genetic research. In the context of Cre AAV2, the virus is designed to carry Cre recombinase, which induces recombination at specific loxP sites within the host genome. This enables researchers to activate or deactivate specific genes in a controlled manner, facilitating the study of gene function and modeling of disease.

Despite the advantages of the Cre/loxP recombination system in animal models, its application in rats has lagged behind that in mice due to the lack of relevant reporter strains. Here, researchers generated a floxed STOP tdTomato rat that conditionally expresses a red fluorescent protein variant (tdTomato) in the presence of exogenous Cre recombinase. The tdTomato signal vividly visualized neurons, including their projection fibers and spines, without any histological enhancement. In addition, a transgenic rat line (FLAME) that ubiquitously expresses tdTomato was successfully established by injecting intracytoplasmic Cre mRNA into fertilized eggs. The rat reporter system here will facilitate connectome studies and visualization of the fine structures of genetically identified cells for extended periods of time in vivo and in vitro. In addition, FLAME is an ideal model for organ transplantation studies due to the improved traceability of cells/tissues.

Here, to examine tdTomato expression after complete excision of the STOP cassette filler, AAV2-Cre virus vectors containing Cre was injected into the striatum (n = 8), hippocampus (n = 7), and cerebellum (n = 2) of heterozygous rats obtained by crossing male NBRP-0734 with wild-type female LE rats. In all rats, bright red fluorescence was detected only in the area surrounding the injection site. Cre-immunoreactive nuclei (Alexa Fluor 488) and tdTomato immunoreactivity (Alexa Fluor 546) colocalized within the same cells (Figure 1A). Almost all cells expressing Cre were successfully recombined (Figure 1B). Even after paraformaldehyde fixation, tdTomato fluorescence signals were retained, and fine structures such as spines could still be resolved without immunohistochemical enhancement (Figures 1C and 1D). Upon closer inspection, tdTomato was expressed primarily in neurons, presumably the neuronal target of the virus serotype. The fluorescence from the projection fibers was visible even without histological enhancement.

AAV2-Cre-mediated reporter gene expression in the brain.Figure 1. AAV2-Cre-mediated reporter gene expression in the brain. (Igarashi H, et al., 2016)

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Customer Reviews
Easy to Handle

The protocols are straightforward, and the support materials provided are comprehensive, making it accessible even for those new to viral vector handling.

Canada

11/05/2021

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