Multiple-gene editing system using a non-inheritable maternal Cas9 in mice
| Project/Area Number |
26670153
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| Research Category |
Grant-in-Aid for Challenging Exploratory Research
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| Allocation Type | Multi-year Fund |
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| Research Field |
Pathological medical chemistry
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| Research Institution | Shinshu University |
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Principal Investigator |
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| Co-Investigator(Kenkyū-buntansha) |
SHINDO Takayuki 信州大学, 学術研究院医学系, 教授 (90345215)
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| Project Period (FY) |
2014-04-01 – 2016-03-31
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| Project Status |
Completed (Fiscal Year 2015)
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| Budget Amount *help |
¥3,640,000 (Direct Cost: ¥2,800,000、Indirect Cost: ¥840,000)
Fiscal Year 2015: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
Fiscal Year 2014: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
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| Keywords | 疾患モデル動物 / 遺伝子改変マウス / CRISPR / Cas9 / ゲノム編集 / 発生工学 / アドレノメデュリン / RAMP / 受容体修飾因子 / 血管作動性ペプチド / CRISPR/Cas9 / 血管作動性ペプチト |
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| Outline of Final Research Achievements |
The aim of this study was to develop new strategies to generate multiple-gene edited mice by using CRISPR/Cas9 system. First, we established a single blastocyst-based assay for rapidly determining whether CRISPR/Cas9-mediated genome editing worked in mouse preimplantation embryos. Then, we established transgenic (Tg) mice with systemic Cas9 overexpression (sCAT) to use non-inheritable maternal Cas9 (maCas9) protein derived from the zygotes. The maCas9 protein in zygotes derived from mating or in vitro fertilization of Tg/+ oocytes and +/+ sperm could successfully edit the target genome, and modified nine target genes simultaneously after injection with nine different gRNAs alone. Furthermore, we demonstrated the creation of “Cas9 transgene-free” gene-edited mice using non-Tg (+/+) zygotes carrying maCas9. These results suggest that sCAT zygotes have the potential to create multiple gene-modified mice both with and without the Cas9 transgene, enabling repeatable genome engineering.
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Report
(3 results)
Research Products
(26 results)
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[Presentation] Pathophysiological Function of Endogenous Calcitonin Gene-related Peptide in Ocular Vascular Diseases2015
Author(s)
Yuichi Toriyama, Yasuhiro Isesato, Akira Imai, Takayuki Sakurai, Akiko Kamiyoshi, Yuka Ichikawa-Shindo, Hisaka Kawate, Akihiro Yamauchi, Kyoko Igarashi, Megumu Tanaka, Tian Liu, Xian Xian, Liuyu Zhai, Shinji Owa, Toshinori Murata, Takayuki Shindo
Organizer
第79回日本循環器学会
Place of Presentation
大阪国際会議場 (大阪府大阪市)
Year and Date
2015-04-24
Related Report
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[Presentation] Adrenomedullin-RAMP2 system regulates lipid and energy metabolism in white and brown adipocytes2014
Author(s)
Kamiyoshi A, Sakurai T, Ichikawa-Shindo Y, Kawate H, Uetake R, Yamauchi A, Igarashi K, Toriyama Y, Tanaka M, Imai A, Liu T, Xian X, Shindo T.
Organizer
The 18th International Vascular Biology Meeting
Place of Presentation
京都国際会館 (京都)
Year and Date
2014-04-14 – 2014-04-17
Related Report
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