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.

 

PROPRIETARY AAV PLASMID SYNTHESIS WORKFLOW

FEATURES & BENEFITS

  • Industry-Leading Turnaround Time
    And competitive price to save your budget

  • Ph.D.-level Technical Support
    Throughout your project

  • Extensive experience working with a wide variety of custom AAV vectors with ITR synthesis capability

  • FREE codon optimization tools based on an advanced algorithm for high protein expression

  • 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

  • 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

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