AAV-Mediated Gene Delivery to Mouse Brown Adipose Tissue.

ABSTRACT

Recombinant adeno-associated virus (AAV) vectors are attractive vehicles for gene therapy. Yet targeting adipose tissue is still a challenging task. We recently showed that a novel engineered hybrid serotype Rec2 displays high efficacy of gene transfer to both brown and white fat. Furthermore, the administration route influences the tropism and efficacy of Rec2 vector with oral administration transducing interscapular brown fat, while intraperitoneal injection preferentially targets visceral fat and liver. To restrict off-target transgene expression in the liver, we further develop a single rAAV vector harboring two expression cassettes one using CBA promoter driving a transgene and another using a liver-specific albumin promoter driving a microRNA targeting the woodchuck posttranscriptional regulatory element (WPRE) sequence in this rAAV vector. In vivo studies by our lab and others have shown that the Rec2/dual-cassette vector system provides a powerful tool for gain-of-function and loss-of-function studies. Here we offer an updated protocol for AAV packaging and delivery to brown fat.