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article download pdf lna-modified oligonucleotide probes mir-122a-regulated expression cag-promoter-based sensor secreted luciferase sensor exogenous mir-34a activity drug-induced liver injury china life sci secreted gaussia luciferase life sciences article wang xiaobing wu long-term monitoring gaussia princeps luciferase open access privacy choices/manage cookies vertebrate microrna expression developmentally regulated patterns virus rna abundance virus core protein yue wang regulate transgene expression driving transgene expression vivo microrna activity elucidating mirna biology hydrodynamics-based transfection hydrodynamic gene transfer microrna recognition elements main content log mir-34a activities dualluciferase reporter system search search article cite suicide gene therapy cag promoter rendered tissue-specific microrna 1093/nar/gkm1080 mol imaging biol cmv-promoter silencing liver-specific microrna european economic area full access post-transcriptional modifications nucleic acids res papillary thyroid carcinoma environ health perspect altering antitumor effects wild-type adenovirus monitoring tumor progression secreted luciferase

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         headline:Long-term ex vivo monitoring of in vivo microRNA activity in liver using a secreted luciferase sensor
         description:Technology for monitoring in vivo microRNA (miRNA) activity is extremely important for elucidating miRNA biology. However, in vivo studies of miRNA have been hampered by the lack of a convenient approach to reliably reflect real-time functional changes in miRNAs. Sensors for miRNA were developed by adding miRNA target sequences to the 3′-untranslated region of Gaussia princeps luciferase (Gluc) mRNA. These sensors were then evaluated in vitro and in vivo by measuring Gluc activity in cell supernatants and in peripheral blood. Sensors driven by the CMV promoter were effective for monitoring miR-122 in living cells, but not for the long-term monitoring of miR-122 or miR-142 in mouse liver because of CMV-promoter silencing. Replacing the CMV promoter with a CAG promoter rendered these sensors effective for the long-term monitoring of relevant liver miRNA activities. We subsequently used the CAG-promoter-based sensor for the long-term monitoring of endogenous liver miR-122, miR142 and miR-34a activities, as well as for exogenous miR-34a activity. Our study demonstrates that real-time in vivo activities of miRNAs can be continuously and conveniently detected in mouse liver using the sensors that we have developed.
         datePublished:2011-05-15T00:00:00Z
         dateModified:2011-05-15T00:00:00Z
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      headline:Long-term ex vivo monitoring of in vivo microRNA activity in liver using a secreted luciferase sensor
      description:Technology for monitoring in vivo microRNA (miRNA) activity is extremely important for elucidating miRNA biology. However, in vivo studies of miRNA have been hampered by the lack of a convenient approach to reliably reflect real-time functional changes in miRNAs. Sensors for miRNA were developed by adding miRNA target sequences to the 3′-untranslated region of Gaussia princeps luciferase (Gluc) mRNA. These sensors were then evaluated in vitro and in vivo by measuring Gluc activity in cell supernatants and in peripheral blood. Sensors driven by the CMV promoter were effective for monitoring miR-122 in living cells, but not for the long-term monitoring of miR-122 or miR-142 in mouse liver because of CMV-promoter silencing. Replacing the CMV promoter with a CAG promoter rendered these sensors effective for the long-term monitoring of relevant liver miRNA activities. We subsequently used the CAG-promoter-based sensor for the long-term monitoring of endogenous liver miR-122, miR142 and miR-34a activities, as well as for exogenous miR-34a activity. Our study demonstrates that real-time in vivo activities of miRNAs can be continuously and conveniently detected in mouse liver using the sensors that we have developed.
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