RNA interfering approach for clarifying the PPARgamma pathway using lentiviral vector expressing short hairpin RNA.
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.
Department of Biopharmaceutics, Graduate School of Pharmaceutical Science, Osaka University, Osaka 565-0871, Japan.
Kazufumi
K
Katayama
Koichiro
K
Wada
Hiroyuki
H
Miyoshi
Kozo
K
Ohashi
Masashi
M
Tachibana
Rie
R
Furuki
Hiroyuki
H
Mizuguchi
Takao
T
Hayakawa
Atsushi
A
Nakajima
Takashi
T
Kadowaki
Yasuo
Y
Tsutsumi
Shinsaku
S
Nakagawa
Yoshinori
Y
Kamisaki
Tadanori
T
Mayumi
D041721:3T3-L1 Cells; D017667:Adipocytes / Q000166:cytology / Q000378:metabolism*; D000818:Animals; D015153:Blotting, Western; D002454:Cell Differentiation; D002455:Cell Division; D002472:Cell Transformation, Viral; D005819:Genetic Markers; D005822:Genetic Vectors; D016086:Lentivirus / Q000235:genetics*; D008055:Lipids / Q000096:biosynthesis; D051379:Mice; D035683:MicroRNAs / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000032:analysis; D018160:Receptors, Cytoplasmic and Nuclear / Q000037:antagonists & inhibitors / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013997:Time Factors; D014157:Transcription Factors / Q000037:antagonists & inhibitors / Q000378:metabolism*
D005819:Genetic Markers; D008055:Lipids; D035683:MicroRNAs; D012333:RNA, Messenger; D018160:Receptors, Cytoplasmic and Nuclear; D014157:Transcription Factors