Seattle Genova offers outstanding quality AAV packaging services to benefit your AAV-based gene therapy experiments. We have improved a series of proprietary technologies and reagents that have greatly improved recombinant AAV production protocols in terms of titer, purity, potency and consistency, particularly for the AAV vector systems employed in our vector cloning services. As an outcome, we have a growing base of highly satisfied clients who come back to us time after time for their AAV vector cloning and AAV packaging requirements.
Skeletal muscle is an interesting target for gene delivery because of its mass and because the vectors can be provided in a noninvasive way. Adeno-associated virus (AAV) vectors are eligible of transducing skeletal muscle fibers and attaining stable and safe transgene expression. To date, greatly animal experiments utilizing AAV have been based on AAV serotype 2, but some recent researches have indicated that AAV1 is more productive than AAV2/2 in transducing muscle fibers. Newly, novel AAVs (AAV7 and AAV8) were separated from rhesus macaques.
The adeno-associated virus (AAV) vector is a productive device for gene delivery in skeletal muscle. AAV-based therapies indicate promising outcomes for treatment of several genetic disorders, containing muscular dystrophy. These dystrophies illustrate a heterogeneous group of diseases affecting muscles and generally characterized by progressive skeletal muscle wasting and weakness and the improvement of fibrosis. The tropism of each AAV serotype has been greatly studied utilizing systemic delivery routes, but very few researches have compared their transduction efficiency through direct intramuscular injection. Yet, in some muscular dystrophies, where only a few muscles are mainly affected, a local intramuscular injection to target these muscles would be the greatly appropriate route.
The findings show that AAV2/7 and AAV2/8 are able to transduce muscle fibres of immunocompetent mice very efficiently, giving new perspectives in the gene transfer of skeletal muscle.
Also, rAAV6 vectors are eligible of transducing the diaphragm and intercostal muscles of mice after a simple injection into the intrathoracic cavity and are eligible of widespread transduction throughout the musculature of mice inserted in the intraperitoneal space as newborn pups.
Suitable serotypes for skeletal muscle
AAV1, AAV6, AAV7,AAV8,AAV9
We begin by subcloning the gene of interest (GOI), shRNA or gRNA into an associated pAAV cis-plasmid.
Large-scale Production (Optional)
Large-scale preparation of the pAAV cis-plasmid and complimentary plasmids utilizing Qiagen Endo-free Mega Prep kits.
Large scale Transfection
Large-scale transfection of engaged plasmids into 40x15cm plates of HEK293 cells.
Harvest the AAV production cells and purify the AAVs through a sequence of CsCl centrifugations.
Find the titer of the viral stock (in genome copy number per ml, or GC/ml) through quantitative real-time PCR.
Genome-wide AAV Products
Key Features and Benefits
•Ready to utilize for in vitro and in vivo applications
•High titer: improved transduction efficiency
•Transduction of dividing and non-dividing cells
•Expert Technical Support
•Quality and user friendly
1.Mercuri E, Bönnemann CG, Muntoni F. Muscular dystrophies. Lancet. (2019) 394:2025–38.
2.Dan Wang, Alexander Brown, Guangping Gao, Viral Vectors for Muscle Gene Therapy, Muscle Gene Therapy, 10.1007/978-3-030-03095-7, (179-192), (2019).
3.Robin Duelen, Domiziana Costamagna, Maurilio Sampaolesi, Stem Cell Therapy in Muscle Degeneration, The Plasticity of Skeletal Muscle, 10.1007/978-981-10-3292-9, (55-91), (2017).
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