RNAi-based gene silencing in primary mouse and human adipose tissues
Authors
Puri, VishwajeetChakladar, Abhijit
Virbasius, Joseph V.
Konda, Silvana
Powelka, Aimee M.
Chouinard, My T.
Hagan, G. Nana
Perugini, Richard A.
Czech, Michael P.
Document Type
Journal ArticlePublication Date
2006-11-10Keywords
Adaptor Proteins, Signal TransducingAdipose Tissue, White
Animals
Cells, Cultured
Gene Expression Regulation
Glucose
Glucose Transporter Type 1
Glucose Transporter Type 4
Humans
Male
Mice
Nuclear Proteins
PTEN Phosphohydrolase
Phosphorylation
Proto-Oncogene Proteins c-akt
*RNA Interference
RNA, Messenger
Up-Regulation
Life Sciences
Medicine and Health Sciences
Metadata
Show full item recordAbstract
Cultured adipocyte cell lines are a model system widely used to study adipose function, but they exhibit significant physiological differences compared with primary cells from adipose tissue. Here we report short interfering RNA-based methodology to selectively attenuate gene expression in mouse and human primary adipose tissues as a means of rapidly validating findings made in cultured adipocyte cell lines. The method is exemplified by depletion of the PTEN phosphatase in white adipose tissue (WAT) from mouse and humans, which increases Akt phosphorylation as expected. This technology is also shown to silence genes in mouse brown adipose tissue. Previous work revealed upregulation of the mitochondrial protein UCP1 in adipose cells from mice lacking the gene for the transcriptional corepressor RIP140, whereas in cultured adipocytes, loss of RIP140 has a little effect on UCP1 expression. Application of our method to deplete RIP140 in primary mouse WAT elicited markedly increased oxygen consumption and expression of UCP1 that exactly mimics the phenotype observed in RIP140-null mice. This ex-vivo method of gene silencing should be useful in rapid validation studies as well as in addressing the depot- and species-specific functions of genes in adipose biology.Source
J Lipid Res. 2007 Feb;48(2):465-71. Epub 2006 Nov 8. Link to article on publisher's siteDOI
10.1194/jlr.D600033-JLR200Permanent Link to this Item
http://hdl.handle.net/20.500.14038/38280PubMed ID
17093294Related Resources
Link to Article in PubMedae974a485f413a2113503eed53cd6c53
10.1194/jlr.D600033-JLR200