AAV Packaging
GENEWIZ AAV Packaging service centers around our proprietary AAV Vector Production process to provide industry-leading project quality, completion time, and flexibility. The adeno-associated virus (AAV) is a powerful vehicle for cell and 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 the replication and encapsidation of viral DNA. Our support for AAV-based research and therapeutic applications enables us to deliver results for any AAV challenge.
The AAV genome is packaged by using two copies of inverted terminal repeats (ITR) to flank the expression cassette and facilitate vector DNA replication as single-stranded DNA (ssDNA) and packaging into empty preformed capsids. Small rep proteins are needed to assist in the translocation of the AAV genome into the capsid.
What is the packaging limit for AAV?
Adeno associated viruses (AAVs) have a packaging limit of roughly 4.7kb, which is approximately half the packaging limit of vectors from lentiviruses and adenoviruses.
AAV Packaging Applications
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Therapeutic Development: Treat diseases of protein expression and cellular function without gene editing
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CRISPR/Gene Editing: Create targeting mRNA sequences for delivery of CRISPR/Cas9 and other gene editing platforms
AAV Packaging Workflow
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.
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1. AAV ITR Sequencing
Preservation of ITR Integrity: GENEWIZ breakthrough Sanger Sequencing and ITR correction process reads and conserves difficult regions that are critical to AAV success
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2. AAV Plasmid Synthesis
High Fidelity Gene Synthesis: 99.9% reliability in project delivery rate from a team with over 20 years of gene synthesis experience
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3. Cloning and Plasmid Prep
Scalable Viral Prep: Mini- to giga-scale quantities of AAV plasmids available from GENEWIZ proprietary E. coli strains for optimal AAV performance
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4. In Vitro Transcription
AAV Packaging: Generate high-quality viral particles with customizable synthesis, serotype, and scale options
Featured Application: AAV for Therapeutic Delivery
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.
Features and Benefits
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High-Quality Viral Particles – High purity, high yield, with low endotoxin and empty shell
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Rapid Project Completion – AAV packaging in as fast as 13-15 business days (BD) to meet your research deadlines
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Accurate ITR and Transgene Expression – The GENEWIZ proprietary AAV plasmid synthesis workflow ensures better ITR integrity than any other research partner
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Ph.D.–Level Partnership – Our dedicated project management teams provide expert technical consultation to solve any AAV challenge
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Reliable Project Management – Global production facilities and local project management ensures reliable progress and proactive communication
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Customized AAV Delivery – Suit the needs of your project with over 20 serotypes and a wide range of scales for AAV particles
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Total Cell and Gene Therapy Solutions – Protect your IP and reduce logistical delays and errors by working with GENEWIZ as your single partner for sequencing, synthesis, and sample management
AAV Packaging Deliverables
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 Serotypes | Volume | Estimated Completion Time | Quality Control | Titer | Yield |
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| Normal yield: 1, 3b, 5, 7, 8, 9, rh10, PHP.eB, PHP.B, AAVDJ, retro, PHP.S, Olig001, Olig001-PHPA, AAVie | 200 uL 500 uL |
In as few as 3-4 weeks | 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 Custom (up to 100 mL) |
In as few as 3-4 weeks | Low endotoxin (<10 EU/mL) | 1 x 1013 GC/mL | Up to 1 x 1016 GC |
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.
Custom mRNA Synthesis Technical Resources
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Tech Note │ An Efficient and Reliable Approach to AAV Packaging
Previous reports have shown that ITRs are necessary to produce effective viral particles, but their structure is problematic for conventional sequencing, synthesis, and cloning workflows. Here, we study the effect of partial and full ITR deletions on AAV packaging and show how the proprietary methods developed by GENEWIZ can successfully detect and repair these mutations.