title: High-accuracy biodistribution analysis of adeno-associated virus variants by double barcode sequencing.
abstract: Biodistribution analysis is a key step in the evaluation of adeno-associated virus (AAV) capsid variants, whether natural isolates or produced by rational design or directed evolution. Indeed, when screening candidate vectors, accurate knowledge about which tissues are infected and how efficiently is essential. We describe the design, validation, and application of a new vector, pTR-UF50-BC, encoding a bioluminescent protein, a fluorescent protein and a DNA barcode, which can be used to visualize localization of transduction at the organism, organ, tissue, or cellular levels. In addition, by linking capsid variants to different barcoded versions of the vector and amplifying the barcode region from various tissue samples using barcoded primers, biodistribution of viral genomes can be analyzed with high accuracy and efficiency.
authors.affiliation: Department of Pediatrics, Division of Cell & Molecular Therapy, University of Florida , Gainesville, Florida, USA.
authors.forename: Damien
authors.initials: D
authors.lastname: Marsic
authors.affiliation: Department of Molecular Genetics and Microbiology, University of Florida , Gainesville, Florida, USA.
authors.forename: Héctor R
authors.initials: HR
authors.lastname: Méndez-Gómez
authors.affiliation: Department of Pediatrics, Division of Cell & Molecular Therapy, University of Florida , Gainesville, Florida, USA.
authors.forename: Sergei
authors.initials: S
authors.lastname: Zolotukhin