Development of in vitro transcription networks using CRISPR/Cas9 and microfluidic technologies
| Project/Area Number |
17F17796
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| Research Category |
Grant-in-Aid for JSPS Fellows
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| Allocation Type | Single-year Grants |
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| Section | 外国 |
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| Research Field |
Nanobioscience
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| Research Institution | The University of Tokyo |
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Principal Investigator |
藤井 輝夫 東京大学, 生産技術研究所, 教授 (30251474)
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| Co-Investigator(Kenkyū-buntansha) |
BACCOUCHE ALEXANDRE 東京大学, 生産技術研究所, 外国人特別研究員
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| Project Period (FY) |
2017-11-10 – 2020-03-31
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| Project Status |
Completed (Fiscal Year 2019)
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| Budget Amount *help |
¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 2019: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 2018: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 2017: ¥600,000 (Direct Cost: ¥600,000)
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| Keywords | CRISPR/Cas9 / DNA Nanotechnology / Microfluidics / Reaction Network / Fluorescence / fluorescence / microfluidics |
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| Outline of Annual Research Achievements |
In just a few recent years, Cas9 has become an essential tool in the toolbox of molecular biologist. This highly-competitive field holds a lot of potential applications, from fundamental research tools to gene therapy. However, a precise control of its activity, and its incorporation in complex molecular networks have been lacking. The goal of this research project was to gain better control over Cas9 by using tools from DNA nanotechnology and microfluidic. Building on the expertise of the laboratory in molecular programming and microfluidics, new RNA-based mechanisms to control the activity of the enzyme has been developed. The research fellow first adapted a fluorescent assay to quickly and quantitatively assess the activity of a RNA guide. He then tested close to 300 different variants of guides, altering almost any functional region of the guide. During this testing, he found that the nexus, a particular region that forms a short hairpin, was essential to the activity of Cas9. He then demonstrated allosteric modulation of the guide under the control of an exogenous miRNA. Lastly, using this invention, he designed several transcription networks that perform logic operation, such as XOR, AND or OR gates.
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| Research Progress Status |
令和元年度が最終年度であるため、記入しない。
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| Strategy for Future Research Activity |
令和元年度が最終年度であるため、記入しない。
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Report
(3 results)
Research Products
(6 results)
