Development of a novel genome editing-mediated gene knock-in system
| Project/Area Number |
26290070
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Partial Multi-year Fund |
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| Section | 一般 |
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| Research Field |
System genome science
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| Research Institution | Hiroshima University |
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Principal Investigator |
Yamamoto Takashi 広島大学, 理学(系)研究科(研究院), 教授 (90244102)
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| Co-Investigator(Kenkyū-buntansha) |
谷口 俊介 筑波大学, 生命環境科学研究科(系), 准教授 (00505331)
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| Co-Investigator(Renkei-kenkyūsha) |
AIDA TOMOMI 東京医科歯科大学, 難治疾患研究所, 准教授 (50540481)
SAKAMOTO NAOAKI 広島大学, 大学院理学研究科, 准教授 (00332338)
SUZUKI KENICHI 広島大学, 大学院理学研究科, 特任准教授 (90363043)
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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 |
¥17,680,000 (Direct Cost: ¥13,600,000、Indirect Cost: ¥4,080,000)
Fiscal Year 2016: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000)
Fiscal Year 2015: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000)
Fiscal Year 2014: ¥9,360,000 (Direct Cost: ¥7,200,000、Indirect Cost: ¥2,160,000)
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| Keywords | バイオテクノロジー / ゲノム編集 |
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| Outline of Final Research Achievements |
In this study, I established microhomology-mediated end-joining (MMEJ) dependent gene knock-in system that I termed precise integration into target chromosome (PITCh). I performed reporter gene knock-in experiments using PITCh system in cultured cells and animals such as frog and mice. As results, I observed efficient and precise gene knock-in by PITCh method compared with those achieved by HR-mediated gene knock-in. Additionally, I performed genetic screening of MMEJ enhancers and identified that PITCh efficiency upon Exonuclease I (ExoI) overexpression was increased to 2.5 fold compared with that upon mock overexpression. I also confirmed the effect of ExoI overexpression in endogenous gene locus in cultured cells and mouse zygote.
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Report
(4 results)
Research Products
(20 results)
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[Journal Article] Gene cassette knock-in in mammalian cells and zygotes by enhanced MMEJ.2016
Author(s)
Aida T, Nakade S, Sakuma T, Izu Y, Oishi A, Mochida K, Ishikubo H, Usami T, Aizawa H, Yamamoto T, Tanaka K.
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Journal Title
BMC Genomics
Volume: 17
Issue: 1
Pages: 979-979
DOI
Related Report
Peer Reviewed / Open Access / Int'l Joint Research
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[Journal Article] The Expression of TALEN before Fertilization Provides a Rapid Knock-Out Phenotype in Xenopus laevis Founder Embryos.2015
Author(s)
Miyamoto K, Suzuki K, Suzuki M, Sakane Y, Sakuma T, Herberg S, Simeone A, Simpson D, Jullien J, Yamamoto T, Gurdon JB.
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Journal Title
PLOS ONE
Volume: 10(11)
Issue: 11
Pages: e0142946-e0142946
DOI
Related Report
Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
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[Journal Article] Homologous recombination-independent large gene cassette knock-in in CHO cells using TALEN and MMEJ-directed donor plasmids.2015
Author(s)
Sakuma, T., Takenaga, M., Kawabe, Y., Nakamura, T., Kamihira, M. & Yamamoto, T.
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Journal Title
Int. J. Mol. Sci.
Volume: 16
Issue: 10
Pages: 23849-23866
DOI
Related Report
Peer Reviewed / Open Access
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[Presentation] MMEJ-mediated integration of donor DNA in cells and animals using TALENs and CRISPR/Cas92015
Author(s)
Nakade S, Tsubota T, Sakane Y, Kume S, Sakamoto N, Obara M, Daimon T, Sezutsu H, Yamamoto T, Sakuma T and Suzuki KI
Organizer
Keystone Sympsia
Place of Presentation
Big Sky, MT, USA
Year and Date
2015-01-11 – 2015-01-16
Related Report
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