Development of a method to directly introduce a chemically modified DNA cassette into the target region of a genome
| Project/Area Number |
16K07208
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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 |
Genome biology
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| Research Institution | Kazusa DNA Research Institute |
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Principal Investigator |
Nakayama Manabu 公益財団法人かずさDNA研究所, 先端研究開発部, 主任研究員 (30370927)
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| Project Period (FY) |
2016-04-01 – 2019-03-31
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| Project Status |
Completed (Fiscal Year 2018)
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| Budget Amount *help |
¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000)
Fiscal Year 2018: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2017: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2016: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
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| Keywords | バイオテクノロジー / ゲノム改変技術 / 部位特異的組み換え酵素 / VCre/VloxP / SCre/SloxP / エピジェネティックス / 修飾塩基 / ゲノムエンジニアリング / エピジェネティクス / ゲノム / 遺伝子 / 発現制御 / 部位特異的組み換え |
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| Outline of Final Research Achievements |
It is already known that the epigenetic state of a genome is influenced by the epigenetic state of the adjoining region. We developed a novel method to directly introduce a modified DNA cassette into the genome and analyze the influence on its epigenetic state. The foreign gene or chemically modified DNA cassette was effectively introduced into mouse genome by Dual RMCE (recombinase-mediated cassette exchange) using both VCre/VloxP and SCre/SloxP systems. The molecular bar code in the introduced DNA cassette could distinguish the state of each cell by NGS (next generation sequencing) analysis. Moreover, we established VCre- or SCre-expressing mice and their corresponding reporter mice for further analysis using this method.
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| Academic Significance and Societal Importance of the Research Achievements |
我々が独自に開発した新規部位特異的組み換え酵素システム(VCre/VloxPとSCre/SloxP)の発展型としての研究であり、ゲノムツールとしてはユニークな存在である。近年、ゲノム編集ツールのCRISPR/Cas9システムが注目を集めているが、CRISPR/Cas9システムは相同組み換えを利用してノックインを行うために、本研究のようなin vitroで修飾したDNAカセットを直接的にゲノム中に挿入するような手法には用いることはできない。合成した修飾塩基をゲノム中に直接的に導入した後の影響を調べることで修飾塩基の役割とエピジェネティックな状態との関係性がより直接的に明確になるであろう。
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Report
(4 results)
Research Products
(4 results)
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[Journal Article] Novel reporter and deleter mouse strains generated using VCre/VloxP and SCre/SloxP systems, and their system specificity in mice.2018
Author(s)
Yoshimura Y, Ida-Tanaka M, Hiramaki T, Goto M, Kamisako T, Eto T, Yagoto M, Kawai K, Takahashi T, Nakayama M, Ito M.
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Journal Title
Transgenic Res.
Volume: 27
Issue: 2
Pages: 193-201
DOI
Related Report
Peer Reviewed
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