Lentiviral short hairpin ribonucleic acid-mediated knockdown of Akt isoforms in human adipocytes

Objectives: Obesity is associated with severe co-morbidities such as insulin resistance and type 2 diabetes. In some animal models, insulin resistance is associated with an increased rate of fat cell apoptosis. Here, we have generated human insulin resistant adipocytes at the level of Akt using a lentiviral RNAi system in order to study whether insulin resistance interferes with apoptosis sensitivity in human adipocytes. Methods: shRNA sequence templates targeting human Akt1 and Akt2 were designed and cloned into the BLOCK-iT RNAi entry vector. The shRNA cassette was then recombined into a lentiviral vector. Rec…