rAAV-Mediated Gene Delivery to Adipose Tissue

Methods Mol Biol. 2019:1950:389-405. doi: 10.1007/978-1-4939-9139-6_23.

Abstract

Recombinant adeno-associated virus (rAAV) vectors are attractive vehicles for gene therapy. Yet, it is challenging to genetically manipulate adipose tissue in adults due to the low transduction efficiency of naturally occurring AAV serotypes. We recently demonstrated that a novel engineered hybrid serotype Rec2 achieves high transduction of adipose tissue that is superior to naturally occurring serotypes via direct injection to adipose depots. Furthermore, the administration route influences the tropism and efficacy of Rec2 vector: oral administration transduces interscapular brown fat, while intraperitoneal injection preferentially targets visceral fat. Multiple in vivo studies by our lab and others have demonstrated that Rec2 vector provides a powerful tool to genetically manipulate adipose tissue for basic research and potential gene therapies of genetic and acquired diseases. Here we provide detailed protocols for AAV production and delivery to adipose tissue by direct injection, oral administration, and intraperitoneal injection.

Keywords: Brown fat; Direct injection; Intraperitoneal administration; Oral administration; Subcutaneous fat; Visceral fat; rAAV serotype Rec2.

MeSH terms

  • Adipose Tissue / metabolism*
  • Adipose Tissue, Brown / metabolism
  • Animals
  • Dependovirus / genetics*
  • Dependovirus / isolation & purification
  • Gene Transfer Techniques*
  • Genetic Vectors / administration & dosage
  • Genetic Vectors / genetics*
  • Genetic Vectors / isolation & purification
  • Injections, Intraperitoneal
  • Mice