Adeno-Associated Vector (AAV) Information

Adeno-Associated Vector (AAV) Information : Adeno-associated viruses (AAVs) are nonenveloped viruses that package a linear, single-stranded DNA (ssDNA) genome of approximately 4.7 kb. Their lack of pathogenicity, their low capacity to induce cellular immune response, their ability to infect both dividing and nondividing cells, as well as their long-term gene expression, have made them valuable tools for in vitro and in vivo gene delivery.

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– Tropism: wide, dependent on serotype.

– Common Applications: medium to long term in vivo studies.

– Procedures:
– Cloning of the expression cassette in an ITR-containing plasmid.
– Triple transfection of HEK-293 cells (standard production: 4x10E8 cells).
– Standard vector purification: PEG precipitation / iodixanol gradient.

– Characterization:
– Quantification of genome containing particles (gc).

– Quality Control:
– Sterility tests: Tryptic Soy Broth and Fluid Thioglycollated Medium prepared according to the recommendations of the current European and United States Pharmacopeia.
– Mycoplasm test.

– Average total yields: 1013 gc

– Average titers: 7×1012 gc/mL

– Production time of AAV Vector (without cloning): 1-1,5 months.

– Additional Advantage: Pseudotyping allows the packaging of a given expression cassette within any given serotype capsid.

The tropism of each AAV serotype is indicated in the chart below.

Tissue Optimal Serotype
CNS AAV1, AAV2, AAV8, AAV9, pHp.eB, rh10
Eye AAV1, AAV2, AAV4, AAV5, AAV8, rh10
Heart AAV1, AAV6, AAV8, AAV9, DJ, rh10
Kidney AAV2, DJ, rh10
Liver AAV8, AAV9, DJ, rh10
Lung AAV5, AAV6, AAV9, rh10
Pancreas AAV8, AAV9, rh10
Skeletal Muscle AAV1, AAV6, AAV7, AAV8, AAV9
Cross blood brain barrier 9P31
Muscle myoAAV, rh10
Spinal Nerves Retro
Spleen DJ