Development of novel gene therapy strategy by insertion of artificial miRNA sequence into intronic region of endogenous gene(Fostering Joint International Research)
| Project/Area Number |
16KK0189
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| Research Category |
Fund for the Promotion of Joint International Research (Fostering Joint International Research)
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| Allocation Type | Multi-year Fund |
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| Research Field |
Laboratory animal science
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| Research Institution | Tokai University |
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Principal Investigator |
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| Project Period (FY) |
2017 – 2019
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| Project Status |
Completed (Fiscal Year 2019)
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| Budget Amount *help |
¥10,660,000 (Direct Cost: ¥8,200,000、Indirect Cost: ¥2,460,000)
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| Keywords | 人工miRNA / 遺伝子治療 / ハイドロダイナミクス法 / ゲノム編集 / イントロン / 生活習慣病 / ノックダウン / インスリン抵抗性 |
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| Outline of Final Research Achievements |
The CRISPR genome editing system can be a powerful strategy for gene therapy approach. I have been trying to develop a new system to regulate gene expression by CRISPR-assisted insertion of artificial miRNA sequence into an intronic region of an endogenous gene and to apply this method for future gene therapy of lifestyle-related disease. The proof-of-concept study was performed by generating new mouse model in which diabetes-related gene was knocked down in liver cells to improve impaired glucose tolerance. The mouse developed could be a novel animal model for impaired glucose tolerance research. This study also optimized hydrodynamic gene delivery method and showed that hydrodynamic delivery of ribonucleoprotein (RNP) with Cas9 protein mixed with synthetic gRNA elicit better genome editing efficiencies than the plasmid vector-based system in mouse liver.
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| Academic Significance and Societal Importance of the Research Achievements |
本研究は、ゲノムDNAの中で通常はあまり使われない領域を人為的に改変することで、疾患に関与する遺伝子の発現をコントロールすると言う新しい方法論を提案するものである。今回、本手法を用いて糖尿病関連遺伝子の肝臓における発現を抑制できることを示し、この独自のコンセプトの実証に成功した。将来的に、本手法の各種生活習慣病治療への応用を目指していきたい。
また、遺伝子治療を考える上で、成体臓器へ目的の試薬を効率良く核酸を送達する方法の改善は重要な課題であるが、今回、ハイドロダイナミクス法を用いたゲノム編集試薬の肝臓への効率の良い送達条件を見出した。遺伝子治療研究に有用な情報を提供できたと言える。
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Report
(2 results)
Research Products
(4 results)
