Gene transfer in the lung using recombinant adeno-associated virus

Curr Protoc Microbiol. 2012 Aug:Chapter 14:Unit14D.2. doi: 10.1002/9780471729259.mc14d02s26.

Abstract

Adeno-associated virus (AAV) is a small replication-deficient DNA virus belonging to the Parvovirinae family. It has a single-stranded ∼4.7-kb genome. Recombinant AAV (rAAV) is created by replacing the viral rep and cap genes with the transgene of interest along with promoter and polyadenylation sequences. The short viral inverted terminal repeats must remain intact for replication and packaging in production, as well as vector genome processing and persistence in the transduction process. The AAV capsid (serotype) determines the tissue tropism of the rAAV vector. In this unit we will discuss serotype selection for lung targeting along with the factors effecting efficient delivery of rAAV vectors to the murine lung. Detailed procedures for lung delivery (intranasal, orotracheal, and surgical tracheal injection), sample collection, and post-mortem tissue processing will be described.

MeSH terms

  • Animals
  • Containment of Biohazards
  • Dependovirus / genetics*
  • Dependovirus / physiology
  • Gene Transfer Techniques*
  • Genetic Vectors / genetics*
  • Genetic Vectors / physiology
  • Immunohistochemistry / methods
  • Lung / virology*
  • Mice
  • Transgenes
  • Virus Replication