| Budget Amount *help |
¥5,330,000 (Direct Cost: ¥4,100,000、Indirect Cost: ¥1,230,000)
Fiscal Year 2013: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2012: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
Fiscal Year 2011: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
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| Research Abstract |
Imaging of endogenous mRNA in the cytoplasm of living cells promises a significant comprehension of post-transcriptional regulation. Fluorescently labeled linear antisense 2'-O-methyl RNA probe can provide a powerful tool in probing endogenous mRNA. Here, we investigated the feasibility of using antisense probes to monitor the dynamic behavior of endogenous cytoplasmic mRNAs. First, rapid hybridization of the probe was confirmed, enabling us to visualize the fast localization of mRNA to stress granules under cellular stress. Next, design of potent antisense probe for a given mRNA target was investigated. Referring to the predicted second structure of mRNA, we have designed and prepared antisense probe candidates. Quantitative analysis of the binding efficiency of these probe based on their intracellular mobility provided some competent antisense probes for GAPDH mRNA. Thus, our approach provides a basis for real time imaging of endogenous cytoplasmic mRNA in living cells.
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