Protocol for Efficient Generation and Characterization of Adeno-Associated Viral Vectors

Hum Gene Ther Methods. 2017 Oct;28(5):235-246. doi: 10.1089/hgtb.2017.192.

Abstract

Adeno-associated virus vectors are a powerful tool for gene transfer approaches. We have established a simple and fast plasmid-based production system for achieving high adeno-associated virus titers within 6 working days. The same procedure can be used for all serotypes and thus allows direct comparability of different serotypes. In this protocol we describe a step-by-step procedure that results in well-characterized vectors suitable for both in vitro approaches and preclinical studies.

Keywords: adeno-associated virus (AAV); quality control; titration; vector purification; viral vector.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Culture Techniques
  • Centrifugation / standards
  • Dependovirus / genetics*
  • Electrophoresis, Polyacrylamide Gel
  • HEK293 Cells
  • Humans
  • Plasmids / analysis
  • Plasmids / metabolism*
  • Plasmids / standards
  • Quality Control
  • Real-Time Polymerase Chain Reaction
  • Transfection