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.
Katayama
Kazufumi
K
Department of Biopharmaceutics, Graduate School of Pharmaceutical Science, Osaka University, Osaka 565-0871, Japan.
Wada
Koichiro
K
Miyoshi
Hiroyuki
H
Ohashi
Kozo
K
Tachibana
Masashi
M
Furuki
Rie
R
Mizuguchi
Hiroyuki
H
Hayakawa
Takao
T
Nakajima
Atsushi
A
Kadowaki
Takashi
T
Tsutsumi
Yasuo
Y
Nakagawa
Shinsaku
S
Kamisaki
Yoshinori
Y
Mayumi
Tadanori
T
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