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JapanesePROPRIETARY AAV PLASMID SYNTHESIS WORKFLOW
AAV PLASMID SYNTHESIS
With over a decade of experience synthesizing DNA, GENEWIZ has developed a unique adeno-associated virus (AAV) plasmid synthesis service, the first of its kind on the market. This service provides researchers working with adeno-associated virus vectors with synthesis and cloning of transgene expression cassettes into custom AAV vectors with high efficiencies. GENEWIZ’s AAV plasmid synthesis service comes bundled with our new AAV plasmid preparation protocol to deliver mini- to giga-scale AAV plasmids that have been sequence-verified by our AAV-ITR Sanger sequencing service.
We understand the importance of intact inverted terminal repeat (ITR) regions in downstream AAV gene therapy applications, as seen in recent publications, therefore GENEWIZ maintains a 2-step QC process and ITR correction by gene synthesis to ensure the integrity of your ITR regions.
FEATURES & BENEFITS
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Industry-Leading Turnaround Time
And competitive price to save your budget -
Ph.D.-level Technical Support
Throughout your project
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Extensive experience working with a wide variety of custom AAV vectors with ITR synthesis capability
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FREE codon optimization tools based on an advanced algorithm for high protein expression
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Ensure intact ITR regions before and after cloning with our robust 2-step QC system including sequence verification using GENEWIZ’s proprietary AAV-ITR sequencing and ITR correction via gene synthesis
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Mini- to giga-scale preps using proprietary AAV plasmid preparation protocol
GENEWIZ INVERTED TERMINAL REPEAT (ITR) CORRECTION
Original AAV inverted terminal repeat shows a deletion mutation. Regular PCR amplification cannot repair the sequence due to its repetitive nature. GENEWIZ can successfully deliver repaired inverted terminal repeat regions through specific ITR correction technique.
IMPORTANCE OF INTACT ITR REGIONS IN AAV GENE THERAPY RESEARCH
Each inverted terminal repeat region is important in downstream AAV gene therapy applications*
| Function | Effecting Region |
| Residue of Non-Recombinant AAV (r(AAV)) Encapsided DNA | A-A’ |
| Packing | A-A’ (trs), D |
| Productivity | B-B’, C-C’ |
| Encapsidation | A-A’, B-B’, C-C’, D |
| Affinity to Rep Protein | A-A’, B-B’, C-C’ |
| Transduction Efficiency | A-A’ |
| Replication | A-A’, D |
| Rescue | A-A’ (trs), D |
| Integration | A-A’ (trs), D |
* Savy, A., Dickx, Y., Nauwynck, L., Bonnin, D., Merten, O.W. and Galibert, L., 2017. Impact of inverted terminal repeat integrity on rAAV8 production using the baculovirus/Sf9 cells system. Human Gene Therapy Methods, 28(5), pp.277-289.
*Zhou, Q., Tian, W., Liu, C., Lian, Z., Dong, X. and Wu, X., 2017. Deletion of the B-B’and C-C’regions of inverted terminal repeats reduces rAAV productivity but increases transgene expression. Scientific reports, 7(1), pp.1-13.
*Maurer, A.C. and Weitzman, M.D., 2020. Adeno-Associated Virus Genome Interactions Important for Vector Production and Transduction. Human Gene Therapy, 31(9-10), pp.499-511.
*trs – terminal resolution site
Technical Resources
Tech Note: An Efficient and Reliable Approach to AAV Packaging
While AAV is a powerful vehicle for gene delivery, unstable ITRs threaten the fidelity of rAAV genomes. Loss of ITR integrity diminishes the packaging efficiency of rAAV particles and thus viral titer. In this tech note, discover the effect of partial and full ITR deletions on AAV packaging and how to reliably identify and effectively correct them.
Tech Note: An Efficient and High-Fidelity Approach to AAV Plasmid Preparation
Azenta has recently developed a proprietary AAV plasmid preparation process, which significantly improves the chance of maintaining ITR integrity, even when other commercially available competent cells growing in low temperature have failed. This process can also isolate clones with full-length ITR from a sample mixture containing intact and truncated ITRs.
Workshop & Roundtable Discussion: A Comprehensive Guide to Using AAV Vectors in Gene Therapy
Working with AAV vectors can be challenging. In this on-demand workshop & roundtable discussion, you’ll gain a better understanding of the AAV development pipeline for gene therapy research and learn how to optimize upstream and downstream processes including AAV synthesis, sequencing, bioinformatics, and storage.
Related Services
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