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dc.contributor.authorPuri, Vishwajeet
dc.contributor.authorChakladar, Abhijit
dc.contributor.authorVirbasius, Joseph V.
dc.contributor.authorKonda, Silvana
dc.contributor.authorPowelka, Aimee M.
dc.contributor.authorChouinard, My T.
dc.contributor.authorHagan, G. Nana
dc.contributor.authorPerugini, Richard A.
dc.contributor.authorCzech, Michael P.
dc.date2022-08-11T08:09:31.000
dc.date.accessioned2022-08-23T16:34:12Z
dc.date.available2022-08-23T16:34:12Z
dc.date.issued2006-11-10
dc.date.submitted2009-03-10
dc.identifier.citationJ Lipid Res. 2007 Feb;48(2):465-71. Epub 2006 Nov 8. <a href="http://dx.doi.org/10.1194/jlr.D600033-JLR200">Link to article on publisher's site</a>
dc.identifier.issn0022-2275 (Print)
dc.identifier.doi10.1194/jlr.D600033-JLR200
dc.identifier.pmid17093294
dc.identifier.urihttp://hdl.handle.net/20.500.14038/38280
dc.description.abstractCultured 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.
dc.language.isoen_US
dc.relation<a href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=pubmed&cmd=Retrieve&list_uids=17093294&dopt=Abstract">Link to Article in PubMed</a>
dc.relation.urlhttp://dx.doi.org/10.1194/jlr.D600033-JLR200
dc.subjectAdaptor Proteins, Signal Transducing
dc.subjectAdipose Tissue, White
dc.subjectAnimals
dc.subjectCells, Cultured
dc.subjectGene Expression Regulation
dc.subjectGlucose
dc.subjectGlucose Transporter Type 1
dc.subjectGlucose Transporter Type 4
dc.subjectHumans
dc.subjectMale
dc.subjectMice
dc.subjectNuclear Proteins
dc.subjectPTEN Phosphohydrolase
dc.subjectPhosphorylation
dc.subjectProto-Oncogene Proteins c-akt
dc.subject*RNA Interference
dc.subjectRNA, Messenger
dc.subjectUp-Regulation
dc.subjectLife Sciences
dc.subjectMedicine and Health Sciences
dc.titleRNAi-based gene silencing in primary mouse and human adipose tissues
dc.typeJournal Article
dc.source.journaltitleJournal of lipid research
dc.source.volume48
dc.source.issue2
dc.identifier.legacycoverpagehttps://escholarship.umassmed.edu/oapubs/1152
dc.identifier.contextkey770130
html.description.abstract<p>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.</p>
dc.identifier.submissionpathoapubs/1152
dc.contributor.departmentDepartment of Surgery
dc.contributor.departmentProgram in Molecular Medicine
dc.source.pages465-71


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