Adeno-associated Virus (AAV) Technology

AAV vectors and packaging tools for in vivo and non-dividing cell gene delivery

When you need to deliver a transgene to a specific tissue in vivo, or to non-dividing cells where lentivirus does not work, AAV is the routine choice. This section covers AAV packaging plasmids, ready-to-use AAV particles across serotypes (AAV2, 5, 6, 8, 9, PHP.eB), AAV-based CRISPR delivery vectors, and titration controls. Lentivirus, AAV, and adenovirus sit alongside each other: lentivirus for stable integration in dividing cells, AAV for non-dividing or in vivo, adenovirus for high-titre transient.

Five decisions, in order.

1. Serotype. For CNS choose AAV9 or PHP.eB. For muscle choose AAV6 or AAV9. For lung choose AAV5. For liver choose AAV8. For ocular choose AAV2. For unknown target tissue pilot two or three candidates.
2. Payload. AAV packaging is limited to ~4.7 kb cargo. For larger payloads use dual-AAV split systems or switch platform.
3. Titre. For in vivo work need 10^12 to 10^14 GC/mL. Titre by qPCR; UV overestimates infectious titre by 3- to 10-fold.
4. Purity. For in vivo studies choose iodixanol-gradient-purified or HPLC-purified. Impurities cause inflammatory responses.
5. Ready-to-use versus packaging plasmid. For one-off in vivo studies use ready-to-use. For sustained in-house production use packaging plasmid systems.

For packaging plasmid systems, GeneCopoeia. For titration and characterisation tools, Cell Biolabs. For AAV purification reagents, Norgen Biotek and AkrivisBio.