| Project/Area Number |
24592806
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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 |
Functional basic dentistry
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| Research Institution | Saga University |
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Principal Investigator |
Kukita Akiko 佐賀大学, 医学部, 准教授 (30153266)
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| Co-Investigator(Kenkyū-buntansha) |
Kukita Toshio 九州大学, 大学院歯学研究院, 教授 (70150464)
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| Co-Investigator(Renkei-kenkyūsha) |
Nishioka Kenichi 佐賀大学, 医学部, 講師 (80370120)
Kuhara Satoru 九州大学, 大学院農学研究院, 教授 (00153320)
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| Project Period (FY) |
2012-04-01 – 2016-03-31
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| Project Status |
Completed (Fiscal Year 2015)
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| Budget Amount *help |
¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000)
Fiscal Year 2014: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2013: ¥2,470,000 (Direct Cost: ¥1,900,000、Indirect Cost: ¥570,000)
Fiscal Year 2012: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
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| Keywords | 破骨細胞分化 / shRNAライブラリー / スクリーニング / 破骨細胞 / shRNA / 分化 / 細胞分化 / 膜タンパク質 / RANKL / マクロファージ / レポーター細胞 |
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| Outline of Final Research Achievements |
We performed the screening for proteins which have essential roles in osteoclast differentiation and function by using shRNA lentivirus library that target signaling pathway in mouse. First, we prepared a reporter cell line RAW-ctsk-Venus1 which induce the expression of fluorescent protein Venus under the control of cathepsin K promoter. RANKL dose dependently induced Venus expression in RAW-ctsk-Venus1. Surface expression of population of Venus-positive cells was analyzed. RANK expression of Venus low-positive cells was lower than that of Venus high-positive cells. RAW-ctsk-Venus 1 cells were infected by shRNA lentivirus library and stimulated with RANKL. The fraction of Venus low-positive cells affected by infection were collected by cell sorter. Analyzing of enriched population of shRNA in Venus low-positive cells revealed that several candidates for regulators of osteoclastogenesis including some signaling molecules Map3k and Vav3.
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