As cell and gene therapy research continues to advance across a wide-range of diseases, the need for effective vectors for successful therapy delivery has increased. Identified as an optimal vehicle, adeno-associated virus (AAV) plasmids are the leading viral vector used in in vivo gene therapy clinical trials due to their high efficiency and enhanced safety in humans.
As a global leader in genomics services, GENEWIZ has developed unique AAV-ITR sequencing, AAV plasmid synthesis, and AAV plasmid preparation capabilities to support scientists in their in vivo gene therapy research. GENEWIZ’s ground-breaking AAV services also include sequence verification and correction of inverted terminal repeat (ITR) regions, which are crucial for rAAV packaging production.
Utilizes Sanger protocols to sequence-confirm difficult ITR regions, expediting screening and validation of lead candidates. Whole AAV plasmid sequencing through primer walking is also available.
Enables synthesis and cloning of transgene expression cassettes into custom AAV vectors. Includes ITR correction to synthesize and clone correct ITR sequences.
Adeno-associated virus (AAV) is one of the most actively investigated gene therapy vehicles. AAV has a linear single-stranded DNA (ssDNA) genome of approximately 4.7 kilobases (kb), with two 145 nucleotide-long inverted terminal repeats (ITR) at the termini which are crucial for rAAV production.
Figure 1. An AAV transfer plasmid containing the modified/transgene expression cassette with intact ITRs is generated and cloned into a plasmid along with Rep/Cap and helper plasmids, which are co-transfected into cell lines for virus packaging and rAAV production.
GENEWIZ has developed a high-quality, direct Sanger sequencing method that reads through both intact and commonly mutated inverted terminal repeat (ITR) regions of adeno-associated virus (AAV). This method is an effective tool for assessing the integrity of ITRs in AAV plasmids.