Adeno-Associated Vector (AAV) Information

Adeno-Associated Vector (AAV) Information : These nonenveloped viruses 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.

– 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).
– Infectivity tests not included.

– 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: 1,5×10¹³ gc

– Average titers: 10¹³ 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
iPSC	AAV7m8
Cross blood brain barrier	9P31, VCAP-102
Muscle	myoAAV, rh10
Spinal Nerves	Retro
Spleen	DJ
PNS	pHp.S