Genetic humanized Mouse production by CRISPITT
Project/Area Number |
26870077
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Research Category |
Grant-in-Aid for Young Scientists (B)
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Allocation Type | Multi-year Fund |
Research Field |
Integrative animal science
Laboratory animal science
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Research Institution | University of Tsukuba |
Principal Investigator |
MIZUNO Seiya 筑波大学, 医学医療系, 助教 (10633141)
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Research Collaborator |
Sugiyama Fumihiro 筑波大学, 医学医療系, 教授 (90226481)
Takahashi Takeshi 実験動物中央研究所, 免疫研究室, 室長 (80335215)
Iijima Saori
Dinh Thi Huong Tra
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Project Period (FY) |
2014-04-01 – 2017-03-31
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Project Status |
Completed (Fiscal Year 2016)
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Budget Amount *help |
¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000)
Fiscal Year 2016: ¥780,000 (Direct Cost: ¥600,000、Indirect Cost: ¥180,000)
Fiscal Year 2015: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2014: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
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Keywords | ゲノム編集 / 受精卵ゲノム編集 / マウスリソース開発 / ヒト化マウス |
Outline of Final Research Achievements |
In the past, the mice carried targeted gene mutation was produced by gene-targeting method using with embryonic stem cells. This way is reliable but long time and big money were required. Genome editing technology are changing the way to produce the gene targeted mouse. This make it possible to produce the mice carried desired mutation just by embryo-microinjection. Especially, CRISPR/Cas9 is most powerful tool. We can make not only knock-out but also knock-in mouse. To produce knock-in mouse by embryo-based genome editing, single guide RNA, Cas9, and donor DNA were co-microinjected. For short tag (e.g. FLAG, HA. V5) knock-in, single strand oligo DNA donor have beed used. In contrast, double strand DNA vectors were used for gene fragment knock-in. In this study, we validated the several way to produce knock-in by embryo-based genome editing and succeed to produce more than 70 bp knock-in with synthesized single strand DNA and more than 7 kb fragment knock-in with plasmid donor.
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Report
(4 results)
Research Products
(18 results)
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[Journal Article] SHISA6 Confers Resistance to Differentiation-Promoting Wnt/β-Catenin Signaling in Mouse Spermatogenic Stem Cells2017
Author(s)
Moe Tokue, Kanako Ikami, Seiya Mizuno, Chiyo Takagi, Asuka Miyagi14, Ritsuko Takada, Chiyo Noda, Yu Kitadate, Kenshiro Hara, Hiroko Mizuguchi, Takuya Sato, Makoto Mark Taketo, Fumihiro Sugiyama, Takehiko Ogawa, Satoru Kobayashi, Naoto Ueno, Satoru Takahashi, Shinji Takada, Shosei Yoshida
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Journal Title
Stem Cell Reports
Volume: 8
Pages: 561-575
DOI
Related Report
Peer Reviewed / Open Access
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[Journal Article] Forward genetic analysis of sleep in randomly mutagenized mice.2017
Author(s)
Hiromasa Funato, Chika Miyoshi, Tomoyuki Fujiyama, Takeshi Kanda, Makito Sato, Zhiqiang Wang, Jing Ma, Shin Nakane, Jun Tomita, Aya Ikkyu, Miyo Kakizaki, Noriko Hotta-Hirashima, Satomi Kanno, Haruna Komiya, Fuyuki Asano, Takato Honda, Staci J. Kim, Kanako Harano, Hiroki Muramoto, Toshiya Yonezawa, Seiya Mizuno, et. al.
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Journal Title
Nature
Volume: 539
Pages: 378-383
DOI
Related Report
Peer Reviewed / Int'l Joint Research
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[Journal Article] Generation of CRISPR/Cas9-mediated bicistronic knock-in ins1-cre driver mice.2016
Author(s)
Yoshikazu Hasegawa, Yoshikazu Hoshino, Abdelaziz E. Ibrahim, Kanako Kato, Yoko Daitoku, Yoko Tanimoto, Yoshihisa Ikeda, Hisashi Oishi, Satoru Takahashi, Atsushi Yoshiki, Ken-ichi Yagami, Hiroyoshi Iseki, Seiya Mizuno, Fumihiro Sugiyama
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Journal Title
Experimental Animals
Volume: 65
Pages: 319-327
DOI
Related Report
Peer Reviewed
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[Journal Article] Simple generation of albino C57BL/6J mice with G291T mutation in the tyrosinase gene by the CRISPR/Cas9 system.2014
Author(s)
Seiya Mizuno, Tra Thi Huong Dinh, Kanako Kato, Saori Mizuno-Iijima, Yoko Tanimoto, Yoko Daitoku, Yoshikazu Hoshino, Masahito Ikawa, Satoru Takahashi, Fumihiro Sugiyama, Ken-ichi Yagami
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Journal Title
Mammalian Genome
Volume: 25
Pages: 327-334
DOI
Related Report
Peer Reviewed / Acknowledgement Compliant
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[Presentation] Applied Genome Editing with CRISPR/Cas9 Plasmid in Mice2015
Author(s)
Seiya Mizuno, Kanako Kato, Saori Iijima, Yuko Hamada, Tra Dinh Thi Huong, Yoshikazu Hoshino, Yoko Tanimoto, Satoru Takahashi, Fumihiro Sugiyama, Ken-ichi Yagami
Organizer
トランスポゾン転移とゲノム編集技術に関する国際会議2015
Place of Presentation
奈良県新公会堂
Year and Date
2015-11-17 – 2015-11-20
Related Report
Int'l Joint Research
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