Therefore, mice with a uniform genetic background remain the best choice for researchers to conduct HBV research. The rAAV virus vector carries a 1.3×HBV full-length genome, and the mouse model of persistent HBV infection is prepared by one-time tail vein injection. Such method shows advantages include simple preparation, high success rate, uniformity and stability, obvious dose-effect relationship, and wide application range. HBV-AAV has been widely used in HBV drug evaluation and vaccine screening.
This animal model can greatly shorten the preparation time for the preclinical animal experiment stage of hepatitis B drug research and development with a rather low cost, which can greatly accelerate the progress of hepatitis B treatment programs and new drug research. The AAV-HBV viral vectors provided by PackGene Biotech share the significant advantages of good safety, long-term transduction, fast HBV modelling, and stable expression of hepatitis B antigen.
Features of HBV rAAV vector
Compared with Lentivirus and Adenovirus, rAAV shows outstanding safety advantages such as the extremely low probability of genome integration, extremely low immunogenicity, and high experimental operation safety.
In the past 20 years, rAAV has been used as a vector for efficient and long-term gene expression in basic research and clinical gene therapy. According to existing records, its expression in primate muscles can last for more than 10 years.
The capsid proteins of different serotypes of AAV recognize different receptors on the cell surface, so the infection efficiency of cells varies in different tissues, showing a certain organ targeting specificity. Among them, AAV8 is the strongest hepatotropic type among AAV vectors and is often used in liver-related research.
- High purity titer ≥1E+13GC/ml shown regarding AAV8-based qPCR genome copies/ml
- Low endotoxin lower than 10EU/ml, suitable for animal experiments.
- AAV-HBV Mycoplasma test negative.
- Low empty shell: AAV-HBV TEM detects empty shell rate is below 30%.
- Complete quality inspection reports are provided.
AAV Type: AAV8-HBV-0001
Genotype: HBV-D genotype, AYW genotype
Vector: HBV-D-AYW 1.3
Remark: Epidemic worldwide and in part of China
AAV Type: AAV8-HBV-0002
Genotype: HBV-C genotype, ADR genotype
Vector: HBV-C-ADR 1.3
Remark:Mainly epidemic in Asia and China with strong pathogenicity
AAV Type: AAV8-HBV-0003
Genotype: HBV-C2, replication defective
Remark: Epidemic in Asia
AAV Type: AAV8-HBV-0001
Genotype: HBV-B genotype, ADW genotype
Vector: HBV-B-Adw 1.3
Remark: Mainly epidemic in Asia and China with less pathogenicity compared with HBV-C type
In 1994, the safety of rAAV as a gene therapy vector was recognized by the FDA. Biological safety is Grade 1 (BSL-1), which is the same as that of plasmid DNA. It is recommended to apply experiments with BSL-2 laboratory and ClassⅡ biological safety cabinet.
- Store the virus at -80°C, take it out when needed, and place it on ice during operation. Fast Service must be used within 24 hours.
- Calculate the usage in advance, and PackGene will sub-pack the virus according to the predetermined requirements before shipping to avoid re-freezing and thawing sub-packaging after receiving the goods, which might affect the titer accuracy. If sub-packaging is required, it is recommended to use PCR tubes with siliconized inner walls, or special virus preservation tubes with low protein binding rates.
- Melt the virus in an ice bath and dilute with PBS or PBS / 0.001% F-68, if you need to dilute the AAV-HBV.
rAAV has been widely used in animal experiments and cell research. However, please note that, for in vivo experiments, the solutions are various to problems like how to choose the injection site, how much virus to inject, and when to test after injection. It is difficult to have a simple and universal injection solution because of different research fields and animals. It needs to be explored in the experiment.
Design of AAV-HBV Mouse Modeling
Due to individual differences and various operator techniques in model mice, it is strongly recommended that AAV-HBV injection be tested in a gradient of 3 to 4 volumes before the formal experiment, with each injection increasing according to the minimum requirements. For example: a total three serial dilutions: 1E+11GC; 2E+11GC; 3E+11GC.
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