A technology to quantitatively measure protein interactomes
Project/Area Number |
15K18466
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Research Category |
Grant-in-Aid for Young Scientists (B)
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Allocation Type | Multi-year Fund |
Research Field |
System genome science
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Research Institution | The University of Tokyo |
Principal Investigator |
Yachie Nozomu 東京大学, 先端科学技術研究センター, 准教授 (60636801)
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Project Period (FY) |
2015-04-01 – 2018-03-31
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Project Status |
Completed (Fiscal Year 2017)
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Budget Amount *help |
¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000)
Fiscal Year 2017: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2016: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2015: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
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Keywords | DNAバーコード / インタラクトーム / 超並列DNAシークエンシング / 酵母ツーハイブリッド / タンパク質間相互作用 / DNA barcode / タンパク質進化 / 次世代DNAシークエンサー |
Outline of Final Research Achievements |
We remain one or two orders of magnitude away from a complete interaction map for humans and other major model organisms. Completion will require screening at substantially larger scales with many complementary assays, requiring further efficiency gains in proteome-scale interaction mapping. Here, we report Barcode Fusion Genetics-Yeast Two-Hybrid (BFG-Y2H), by which a full matrix of protein pairs can be screened in a single multiplexed strain pool. BFG-Y2H uses Cre recombination to fuse DNA barcodes from distinct plasmids, generating chimeric protein-pair barcodes that can be quantified via next-generation sequencing. We applied BFG-Y2H to four different matrices ranging in scale from ~25 K to 2.5 M protein pairs. The results show that BFG-Y2H increases the efficiency of protein matrix screening, with quality that is on par with state-of-the-art Y2H methods.
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Report
(4 results)
Research Products
(20 results)
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[Journal Article] Systematic analysis of Ca2+ homoeostasis in Saccharomyces cerevisiae based on chemical-genetic interaction profiles2017
Author(s)
Ghanegolmohammadia F, Yoshida M, Ohnuki S, Sukegawa Y, Okada H, Obara K, Kihara A, Suzuki K, Kojima T, Yachie N, Hirata D & Ohya Y
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Journal Title
Molecular Biology of the Cell
Volume: 28
Issue: 9
Pages: 3415-3427
DOI
Related Report
Peer Reviewed
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[Journal Article] Yeast genetic interaction screen of human genes associated with amyotrophic lateral sclerosis: identification of MAP2K5 kinase as a potential drug target2017
Author(s)
Jo M, Chung AY, Yachie N, Seo M, Jeon H, Nam Y, Seo Y, Kim E, Zhong Q, Vidal M, Park HC, Roth FP & Suk K
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Journal Title
Genome Research
Volume: 27
Pages: 1487-1500
Related Report
Peer Reviewed
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[Journal Article] Targeted nucleotide editing using hybrid prokaryotic and vertebrate adaptive immune systems2016
Author(s)
Nishida, K., Arazoe, T., Yachie, N., Banno, S., Kakimoto, M., Tabata, M., Mochizuki, M., Miyabe, A., Araki, M., Hara, K.Y., Shimatani, Z. and Kondo, A.
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Journal Title
Science
Volume: 353
Issue: 6305
Pages: 6305-6305
DOI
Related Report
Peer Reviewed / Open Access / Int'l Joint Research
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[Journal Article] Pooled-matrix protein interaction screens using Barcode Fusion Genetics2016
Author(s)
Yachie N, Petsalaki E, Mellor JC, Weile J, Jacob Y, Verby M, Ozturk SB, Li S, Cote AG, Mosca R, Knapp JJ, Ko M, Yu A, Gebbia M, Sahni N, Yi S, Tyagi T, Sheykhkarimli D, Roth JF, Wong C, Musa L, Snider J, Liu Y-C, Yu H, Braun P, Stagljar I, Hao T, Calderwood MA, Pelletier L, Aloy P, Hill DE, Vidal M & Roth FP
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Journal Title
Molecular Systems Biology
Volume: 12
Issue: 4
DOI
Related Report
Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
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