
Gene therapy is a consistently growing field of research that was experimentally initiated more than 45 years ago for a broad range of diseases, while clinical translation is relatively recent with the first commercial use being approved in 2005 in China. In this aspect, recombinant adeno-associated viral (rAAV) vectors serve as promising tools to induce genetic modifications with low immunogenicity and toxicity, while ensuring the achievement of efficient and long-term transgene expression. They are obtained from the replication-defective adeno-associated virus with basically 12 evident wildtype serotypes that are mainly entering the host cell via heparin sulfate proteoglycan (HSPG) and AAV receptors (AAVR). Recombinant AAV vectors can be either produced using a helper virus (such as adenovirus) or helper-free viruses, where the required adenoviral genes are provided via the helper-plasmid application. Several strategies for rAAV design are available. Naturally occurring single-stranded (ss) AAV vectors require self-priming induced second strand synthesis that leads to delayed transgene expression in the host cell. On the other hand, self-complementary (sc) vectors with dimeric inverted repeat genomes facilitate early transgene expression while the genomic packaging capacity is roughly half-sized. Furthermore, besides covering a broad tissue tropism by the wildtype serotypes, redirection/specific cellular targeting has gained much interest in recent years. For that, natural receptor binding is ablated, and peptide ligands are synthetically fused to the capsid proteins to allow target receptor binding and entry. Hence, methodologies and techniques described herein provide insights into the opportunities of rAAV vector production for different gene therapy approaches/aims.
0 Views
•
Cited by 17
•
2021
•