Adeno-associated viral (AAV) vectors are both a powerful pre-clinical research tool as well as a highly successful clinical gene therapy vehicle. AAV vectors are derived from a small single-stranded DNA virus in the Dependoparvovirus subfamily. The therapeutic gene and regulatory elements are inserted into the vector genome between flanking Inverted Terminal Repeat (ITR) sequences, while replication and structural proteins are provided in trans during the vector production process. High titer production from either mammalian or insect cell lines followed by purification to remove cellular contaminants and vectors lacking complete genome incorporation is imperative in order for successful and reproducible translation of gene therapies on both research and clinical scale. Characterization of vector properties as well as development of in vivo gene therapy models for determining vector efficacy are paramount for development of new therapies.

It is the goal of this Methods Collection to openly share methods in AAV biology and gene therapy development to improve the speed and ease at which the field is able to develop novel life-saving gene therapies. In this collection, we will highlight aspects of AAV-based gene therapy research including vector production and purification, methods of particle characterization, basic AAV biology, and methods in gene therapy model development.