Establishment of a new method for functional analysis of cis-regulatory modules using CRISPRi
| Project/Area Number |
26650087
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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 |
Developmental biology
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| Research Institution | Okinawa Institute of Science and Technology Graduate University |
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Principal Investigator |
Yasuoka Yuuri 沖縄科学技術大学院大学, マリンゲノミックスユニット, 研究員 (70724954)
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| Research Collaborator |
TAIRA Masanori 東京大学, 大学院理学系研究科生物科学専攻, 准教授 (60150083)
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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 |
¥3,120,000 (Direct Cost: ¥2,400,000、Indirect Cost: ¥720,000)
Fiscal Year 2015: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2014: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
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| Keywords | ゲノム編集 / 転写制御 / ツメガエル / 発生 / シス制御領域 / 転写制御関連因子 / 遺伝子発現調節 / 発生ゲノム科学 / CRISPR / CRISPR-Cas9 / 遺伝子発現制御 |
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| Outline of Final Research Achievements |
It is important to analyze in vivo functions of cis-regulatory modules during embryogenesis. However, quick and effective methods for such analysis have never been established. Therefore, I examined whether CRISPRi by a dead Cas9 protein (dCas9) can regulate activities of cis-regulatory modules using Xenopus embryos. Luciferase reporter assays were performed using both dCas9 mRNA and protein. The data showed that CRISPRi could not affect the reporter gene expression. This result suggests that endogenous transcriptional machineries can bump CRISPRi/dCas9 from DNA.
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Report
(4 results)
Research Products
(16 results)
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[Journal Article] Foxh1 Occupies cis-Regulatory Modules Prior to Dynamic Transcription Factor Interactions Controlling the Mesendoderm Gene Program.2017
Author(s)
Rebekah M. Charney, Elmira Forouzmand, Jin Sun Cho, Jessica Cheung, Kitt D. Paraiso, Yuuri Yasuoka, Shuji Takahashi, Masanori Taira, Ira L. Blitz, Xiaohui Xie, and Ken W.Y. Cho.
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Journal Title
Developmental Cell
Volume: 40
Issue: 6
Pages: 595-607
DOI
Related Report
Peer Reviewed / Int'l Joint Research
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[Presentation] Transcriptional basis of the head organizer in the amphibian gastrula: ChIP seq analysis of the head organizer transcription facotors, Otx2, Lim1 and Gsc2014
Author(s)
Yasuoka, Y., Suzuki, Y., Takahashi, S., Someya, H., Sudou, N., Haramoto, Y., Asashima, M., Sugano, S. and Taira, M.
Organizer
第85回日本動物学会年会
Place of Presentation
東北大学川内北キャンパス（宮城県仙台市）
Year and Date
2014-09-12
Related Report
Invited
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