RNA interfering approach for clarifying the PPARgamma pathway using lentiviral vector expressing short hairpin RNA

• Published in: FEBS letters Vol. 560; no. 1-3; pp. 178 - 182
• Main Authors: Katayama, Kazufumi, Wada, Koichiro, Miyoshi, Hiroyuki, Ohashi, Kozo, Tachibana, Masashi, Furuki, Rie, Mizuguchi, Hiroyuki, Hayakawa, Takao, Nakajima, Atsushi, Kadowaki, Takashi, Tsutsumi, Yasuo, Nakagawa, Shinsaku, Kamisaki, Yoshinori, Mayumi, Tadanori
• Format: Journal Article
• Language: English
• Published: England 27.02.2004
• Subjects: 3T3-L1 Cells; Adipocytes - cytology; Adipocytes - metabolism; Animals; Blotting, Western; Cell Differentiation; Cell Division; Cell Transformation, Viral; Genetic Markers; Genetic Vectors; Lentivirus - genetics; Lipids - biosynthesis; Mice; MicroRNAs - metabolism; Promoter Regions, Genetic; Receptors, Cytoplasmic and Nuclear - antagonists & inhibitors; Receptors, Cytoplasmic and Nuclear - metabolism; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger - analysis; Time Factors; Transcription Factors - antagonists & inhibitors; Transcription Factors - metabolism
• ISSN: 0014-5793

Peroxisome proliferator-activated receptor gamma (PPARgamma) plays a central role in adipocyte differentiation and insulin sensitivity. Although PPARgamma also appears to regulate diverse cellular processes in other cell types such as lymphocytes, the detailed mechanisms remain unclear. In this study, we established a lentivirus-mediated short hairpin RNA expression system and identified a potent short hairpin RNA which suppresses PPARgamma expression, resulting in marked inhibition of preadipocyte-to-adipocyte differentiation in 3T3-L1 cells. Our PPARgamma-knockdown method will serve to clarify the PPARgamma pathway in various cell types in vivo and in vitro, and will facilitate the development of therapeutic applications for a variety of diseases.