2012 Fiscal Year Final Research Report
Fluorescent visualization of cancer cellsby monitoring cellular polyamines
| Project/Area Number |
22501019
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Tumor biology
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| Research Institution | Jikei University School of Medicine |
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Principal Investigator |
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| Co-Investigator(Kenkyū-buntansha) |
MURAI Noriyuki 東京慈恵会医科大学, 医学部, 講師 (60300927)
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| Project Period (FY) |
2010 – 2012
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| Keywords | がんの特性 / ポリアミン / 翻訳調節 / 蛍光タンパク質 / アンチザイム |
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| Research Abstract |
We are developing a novel method to visualize cancer cells by combining the polyamine-dependent frameshift mechanism of AZ, an endogenous cellular polyamine sensor, and the fluorescent protein techniques. We created a polyamine sensor construct with frameshift region of AZ1 mRNA flanked by two fluorescent proteins (ECFP and Keima-Red). When the construct was transfected into the cells, however, polyamine-dependent increase of Keima-Red fluorescence was not observed. Therefore, we improved the sensor construct using the entire protein coding region of AZ1 mRNA with EGFP gene that is inserted immediately downstream of the pseudoknot structure. Cells transfected with this construct showedpolyamine-dependent increase of EGFP fluorescence and frameshift product. Further analysis of polyamine response in the cells is continued.
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