There has been persisting research on gene therapy and continues to advance across different diseases. As a result, there has been a rise in effective vectors for the delivery of successful therapy. Known as an optimal vehicle, Adeno-associated virus (AAV) plasmids remain the foremost and leader among other viral vectors used in vivo gene therapy clinical trials. Thanks to their higher efficiency and improved safety in humans. Our addgene aav guide will reveal everything researchers need to know about our custom adeno-associated.
Researchers find the adeno-associated (AAV) as the most suitable and promising candidate when it comes to gene therapy. Although other available vectors are suitable for research works, AAV remains the best among other viral vectors as a result of its non-integrating features. Moreover, in the models of animals, AAVs do not have pathogenicity. They remain a very powerful tool in regenerative medicine. Thanks to their broad tropism as well as their transducing ability to quiescent cells and proliferating cells. Researchers can achieve AAV tissue specificity by using varying AAV serotype capsids. Fortunately, researchers can get different addgene AAV serotypes which fits in when there is a need for Vivo experiments or a crude virus preparation for in vitro cell culture experiments.
Different genomic services are everywhere but we are the global leader among them all. We have developed a unique AAV-ITR sequencing, AAV plasmid synthesis, and AAV plasmid preparation capabilities to support scientists in their in vivo gene therapy research. Our ground-breaking AAV services are available such as verification of sequence and correction of inverted terminal repeat (ITR) regions. These are important when it comes to packaging production of adeno-associated virus (AAV).
What is an adeno-associated virus?
This most sought-after viral vector is a small (20 nm in diameters) replication-defective, nonenveloped virus. Adeno-associated virus (AAV) infects humans as well as some other primate species. At the moment, they are not known to be responsible for causing any kind of disease, and at the same time, they have been shown to cause a very mild immune response. Both dividing and non-dividing cells are not free from being infected by the AAV. Also, AAV may incorporate its genome into the cell that hosts it. This is why we ensure that researchers enjoy every bit of their experiment using our custom adeno-associated virus AAV. With our genetically engineered AAV, we offer custom rAAV services.
Generally, recombinant AAVs are engineered to persist in an extrachromosomal state and could prevent mutation risk. This feature makes AAV lost through division, however, when it comes to non-dividing cells, it can remain stable. So far, AAV expression systems are of two types: Conventional Single-Stranded Adeno-Associated Virus (ssAAV) and Self-Complementary Adeno-Associated Virus (scAAV).
Adeno-associated virus (AAV) Services
- AAV-ITR Sequencing
- Preparation of AAV plasmid preparation
- AAV plasmid synthesis
- AAV genome sequencing
The system of Adeno-associated virus (AAV)
One of the most investigated gene therapy vehicles is the adeno-associated virus (AAV). This vector has a linear single-stranded DNA (ssDNA) genome of about 4.7 kilobases (kb). It also has two 145 nucleotide-long inverted terminal repeats (ITR) at the termini which are crucial for rAAV production.
Advantages of custom AAV services
One of the contributing factors for a researcher to have a successful research outcome is to have the right tools at hand. In the case of experimenting with AAV, achieving success is based on the quality of the vector and titer. Also, it is important to have the correct preferred serotype as well as expression cassette design (such as a promoter). When researchers choose our adeno-associated virus for their experiment it will assist them in identifying the Optima trying for their area of application as well as engineering AAV vectors exactly to their specification to meet their needs. Using our adeno-associated virus AAV services for the AAV expression system comes with lots of advantages. Some of these include:
- It does not integrate into the host gene
- Broad tropism – tissue specificity with various AAV serotype
- There are no elicit significant immune responses in vivo
- There is a long-term expression in non-dividing cells
Why do researchers need AAV services?
- Researchers can use it for the delivery of in vivo gene
- The production is not difficult at a high viral titer ( no helper required)
- Researchers will enjoy personal technical consultation during each stage of their research work
- Topnotch standards as well as upscaling options that will work perfectly from discovery to late preclinical
- Researchers can customize or adjust the process of upstream and downstream steps to work perfectly for the needs of the experiment
- Optimization of expert expression cassette design
- The consistent growth of premium QC selection options that fit into every requirement and experimental needs
- Targeting strong tissue with a state-of-the-art serotype and promoter portfolio
The production process of Custom Adeno-Associated Virus (AAV)
- Subcloning of interest (GOI), shRNA, or gRNA into a related pAAV cis-plasmid is the first thing (not compulsory)
- We prepared pAAV cis-plasmid as well as complimentary plasmids with the use of Qiagen Endo-free Mega Prep kits (not compulsory as well)
- Large-scale transfection of involved plasmids into 40x15cm plates of HEK293 cells.
- Harvest the AAV production cells as well as addgene AAV purification through a series of CsCl centrifugation.
- We determine the viral stock titer (in genome copy number per ml, or GC/ml) through quantitative real-time PCR.
Conclusion
Our custom adeno-associated virus (AAV) is crucial to achieving the best experimental result. We offer top-notch AAV services that fit into all experimental needs and requirements.
