2013 Fiscal Year Final Research Report
Imaging of mRNA in living cells using antisense probe
| Project/Area Number |
23570207
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Research Field |
Molecular biology
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| Research Institution | The University of Tokyo |
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Principal Investigator |
OKABE Kohki 東京大学, 薬学研究科(研究院), 助教 (20455398)
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| Co-Investigator(Renkei-kenkyūsha) |
FUNATSU Takashi 東京大学, 大学院薬学系研究科, 教授 (00190124)
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| Project Period (FY) |
2011 – 2013
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| Keywords | RNA / イメージング / 生細胞 |
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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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