scAAVAnc80L65-CAG-GFP

Different types of viral vectors have been considered for gene delivery in animal models. Replication-defective vectors based on adeno-associated viruses (AAVs) combine robust transduction efficiency with a good in vivo safety profile for many different therapeutic target cells and tissues. Naturally occurring AAVs are not associated with any disease in humans and are able to transduce post-mitotic cells, including neurons. Recombinant AAV vectors used for gene therapy maintain their genome as episomes in non-dividing cells and allow long-term expression. According to the extensive available AAV capsid data, the protein coat protecting the single-stranded DNA genome is the main determinant of its targeting, immunological, and other properties. These properties continue to demonstrate the growing utility of AAVs in clinical gene therapy applications in various fields, as well as the approval of two AAV-based drug products.

Anc80L65 is a novel AAV capsid designed by in silico reconstruction of viral evolutionary lineages. Compared to conventional AAVs, Anc80L65 has previously demonstrated robust transduction capabilities after local delivery in a variety of tissues, such as the liver, retina, or cochlea. Unlike other AAVs, this relatively new vector system was reconstructed based on ancestral sequences and designed in silico in combination with sequence information from natural AAVs, with the primary goal of developing a stable and fully functional AAV variant that is serologically and immunologically distinct from the AAV currently circulating in humans. Importantly, Anc80L65 far outperformed AAV9 in its ability to target the nervous system, and increased the transduction rates of neurons and astrocytes by 3-fold and 4.3-fold, respectively.