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.
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.