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

FEBS Letters

Katayama, Kazufumi²  Mizuguchi, Hiroyuki⁴  Miyoshi, Hiroyuki³  Mayumi, Tadanori²  Ohashi, Kozo²  Wada, Koichiro¹  Hayakawa, Takao⁴  Furuki, Rie²  Tachibana, Masashi²  Kadowaki, Takashi⁵  Kamisaki, Yoshinori¹  Tsutsumi, Yasuo²  Nakagawa, Shinsaku²  Nakajima, Atsushi⁶ 

[1] Department of Pharmacology, Graduate School of Dentistry, Osaka University, 1-8 Yamadaoka, Suita, Osaka 565-0871, Japan;Department of Biopharmaceutics, Graduate School of Pharmaceutical Science, Osaka University, Osaka 565-0871, Japan;Subteam for Manipulation of Cell Fate, BioResource Center, RIKEN, Tsukuba Institute, Ibaraki, Japan;Division of Biological Chemistry and Biologicals, National Institute of Health Sciences, Tokyo 158-8501, Japan;Department of Metabolic Diseases, Graduate School of Medicine, University of Tokyo, Tokyo, 113-0033, Japan;The Third Department of Internal Medicine, Yokohama City University School of Medicine, Yokohama 236-0004, Japan

关键词: Peroxisome proliferator-activated receptor γ;    RNA interference;    Short hairpin RNA;    Lentiviral vector;    Adipocyte;    LV;    lentiviral vector;    shRNA;    short hairpin RNA;    MOI;    multiplicity of infection;    PPAR;    peroxisome proliferator-activated receptor;    GPDH;    glycerol-3-phosphate dehydrogenase;    BRL;    rosiglitazone (BRL-49653);   

DOI  :  10.1016/S0014-5793(04)00100-0

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Peroxisome proliferator-activated receptor γ (PPARγ) plays a central role in adipocyte differentiation and insulin sensitivity. Although PPARγ 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 PPARγ expression, resulting in marked inhibition of preadipocyte-to-adipocyte differentiation in 3T3-L1 cells. Our PPARγ-knockdown method will serve to clarify the PPARγ pathway in various cell types in vivo and in vitro, and will facilitate the development of therapeutic applications for a variety of diseases.

【 授权许可】

Unknown