The adeno-associated virus (AAV) is a powerful vehicle for cell or gene therapy; however, working with AAV vectors can be challenging. Each construct contains two inverted terminal repeat (ITR) sequences which are notoriously unstable, but also critical to replication and encapsidation of viral DNA. To support AAV-based research and therapeutic applications, the GENEWIZ genomics team from Azenta Life Sciences developed an AAV packaging workflow centered around its proprietary AAV plasmid synthesis process for industry-leading project quality, completion time, and flexibility to deliver results for any AAV challenge.
With expertise in Sanger and NGS sequencing, gene synthesis, viral packaging, and more, GENEWIZ has the capabilities and flexibility to partner with you for research success at any stage of your workflow. This means confidence and reliability in your product quality with the convenience of a single team for consultation and service experience. Our total packaging workflow below connects the novel advantages of our research groups to bring you AAV success.
Adeno-associated virus (AAV) is a promising gene therapy vector due to its low pathogenicity and immunogenicity. It can deliver a genetic payload of up to 4.7 kb, and multiple serotypes are available to target specific cell types or tissues in humans. The recombinant AAV (rAAV) genome comprises a transgene construct flanked by a pair of 145 bp inverted terminal repeat (ITR) sequences. Previous reports have shown that the ITRs are necessary to produce viral particles, but their structure is problematic for conventional cloning workflows. ITR sequences in plasmids are unstable, often acquiring deletions during propagation in bacteria. Because of the high risk of mutation, sequence verification of ITR sequences is a critical part of rAAV quality control. This too is challenging, however, as standard Sanger sequencing fails to read through most ITR sequences. Here, we study the effect of partial and full ITR deletions on AAV packaging and show how proprietary methods developed by GENEWIZ can successfully detect and repair these mutations.
AAV particles have a packaging capacity of approximately 4.7 kb and are provided in PBS buffer1. They are shipped on dry ice and aliquoted in custom vialing sizes for convenient storage and use.
|Featured AAV Serotypes2||Volume||Estimated Completion Time||Quality||Titer3||Yield|
|Normal yield: 1, 3b, 5, 7, 8, 9, rh10, PHP.eB, PHP.B, AAVDJ, retro, PHP.S, Olig001, Olig001-PHPA, AAVie||250 uL||
In as few as 13-15 BD
|- Ultrapure (≥95% by SDS-PAGE)||
1 x 1013 GC/mL
Up to 1 x 1016 GC
|Low yield: 2, 4, 6, 6.2, 7m8, AAV ShH10, AAV6 [Y705F,Y731F]||1 mL||- Low endotoxin (<10 EU/mL)|
|Custom (up to 100 mL)|
1. AAV packaging solutions can use GENEWIZ or customer-supplied plasmids.
2. GENEWIZ is continually developing new serotypes through research efforts. Contact our project management team to learn more.
3. Standard quality control includes titer determination by qPCR with ITR primers. Optional dPCR and TEM analysis available.