Development of microinjection-independent animal genome manipulation method
| Project/Area Number |
16H05049
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Integrative animal science
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| Research Institution | National Defense Medical College |
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Principal Investigator |
NAKAMURA Shingo 防衛医科大学校(医学教育部医学科進学課程及び専門課程、動物実験施設、共同利用研究施設、病院並びに防衛, 防衛医学研究センター, 講師 (00505323)
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| Co-Investigator(Kenkyū-buntansha) |
石原 雅之 防衛医科大学校(医学教育部医学科進学課程及び専門課程、動物実験施設、共同利用研究施設、病院並びに防衛, 防衛医学研究センター 医療工学研究部門, 教授 (10508500)
佐藤 正宏 鹿児島大学, 総合科学域総合研究学系, 教授 (30287099)
大塚 正人 東海大学, 医学部, 准教授 (90372945)
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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 |
¥17,160,000 (Direct Cost: ¥13,200,000、Indirect Cost: ¥3,960,000)
Fiscal Year 2018: ¥5,330,000 (Direct Cost: ¥4,100,000、Indirect Cost: ¥1,230,000)
Fiscal Year 2017: ¥5,590,000 (Direct Cost: ¥4,300,000、Indirect Cost: ¥1,290,000)
Fiscal Year 2016: ¥6,240,000 (Direct Cost: ¥4,800,000、Indirect Cost: ¥1,440,000)
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| Keywords | 発生工学 / ゲノム編集 / 遺伝子改変マウス / 遺伝子導入 |
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| Outline of Final Research Achievements |
The traditional transgenic techniques based on microinjection process, require expensive micromanipulator systems and a high level of skill to operate the equipment. In recently, sequence specific nucleases have discovered and revolutionized in the field of developmental engineering. In particular, the CRISPR/Cas9 system has emerged as a very simple and efficient approach to create genome edited mice. Based on such backgrounds, in this study, we attempt to develop microinjection-independent animal genome manipulation method that do not need isolation and ex vivo handling steps but that can deliver the CRISPR/Cas9 system directly in situ. As results, we achieved to elicit CRISPR/Cas9-mediated mutations in a target locus of embryonic cells by intravenous introducing a single plasmid conferring expression of both Cas9 and gRNA via pregnant mice. This may be a useful tool for manipulating the function of embryonic cells in vivo, allowing to create transgenic animal simply.
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| Academic Significance and Societal Importance of the Research Achievements |
遺伝子改変動物の作製では、多くの場合、高価な機材とそれを操作する高度な技術等が必要になる。本研究の目的は、それらを回避できる簡便な新技術を検討することである。その成果は、遺伝子関連の研究を行っている多くの研究者にとって有用なものとなり、特に獣医学、農学、医学分野において貢献できると期待される。また、使用する動物個体数の削減による動物福祉の向上についても期待が持てる。
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Report
(4 results)
Research Products
(27 results)
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[Journal Article] Timing of CRISPR/Cas9-related mRNA microinjection after activation as an important factor affecting genome editing efficiency in porcine oocytes2018
Author(s)
Sato M, Maeda K, Koriyama M, Inada E, Saitoh I, Ohtsuka M, Nakamura S, Sakurai, T, Watanabe S, Miyoshi K
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Journal Title
Theriogenology
Volume: 108
Pages: 29-38
DOI
NAID
Related Report
Peer Reviewed / Open Access
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[Journal Article] The piggyBac-Based Gene Delivery System Can Confer Successful Production of Cloned Porcine Blastocysts with Multigene Constructs.2016
Author(s)
Sato M, Maeda K, Koriyama M, Inada E, Saitoh I, Miura H, Ohtsuka M, Nakamura S, Sakurai T, Watanabe S, Miyoshi K.
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Journal Title
Int J Mol Sci.
Volume: 17
Issue: 9
Pages: 1424-1424
DOI
NAID
Related Report
Peer Reviewed / Open Access
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[Journal Article] Low-molecular weight heparin protamine complex augmented the potential of adipose-derived stromal cells to ameliorate limb ischemia2016
Author(s)
Satoko Kishimoto, Ken-ichi Inoue, Shingo Nakamura, Hidemi Hattori, Masayuki Ishihara, Masashi Sakuma, Shigeru Toyoda, Hideki Iwaguro, Isao Taguchi, Teruo Inoue, Ken-ichiro Yoshida
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Journal Title
Atherosclerosis
Volume: 249
Pages: 132-139
DOI
Related Report
Peer Reviewed / Open Access / Acknowledgement Compliant
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