A novel synthetic-genetic-array–based yeast one-hybrid system for high discovery rate and short processing time
- Chung-Shu Yeh1,6,
- Zhifeng Wang2,6,
- Fang Miao3,
- Hongyan Ma2,
- Chung-Ting Kao3,
- Tzu-Shu Hsu3,4,
- Jhong-He Yu3,
- Er-Tsi Hung3,
- Chia-Chang Lin3,
- Chen-Yu Kuan3,
- Ni-Chiao Tsai3,
- Chenguang Zhou2,
- Guan-Zheng Qu2,
- Jing Jiang2,
- Guifeng Liu2,
- Jack P. Wang2,5,
- Wei Li2,
- Vincent L. Chiang2,5,
- Tien-Hsien Chang1 and
- Ying-Chung Jimmy Lin2,3,5
- 1Genomics Research Center, Academia Sinica, Taipei 11529, Taiwan;
- 2State Key Laboratory of Tree Genetics and Breeding, Northeast Forestry University, Harbin 150040, China;
- 3Department of Life Sciences and Institute of Plant Biology, College of Life Science, National Taiwan University, Taipei 10617, Taiwan;
- 4Institute of Biomedical Informatics and Center for Systems and Synthetic Biology, National Yang-Ming University, Taipei 11221, Taiwan;
- 5Forest Biotechnology Group, Department of Forestry and Environmental Resources, North Carolina State University, Raleigh, North Carolina 27695, USA
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↵6 These authors contributed equally to this work.
Abstract
Eukaryotic gene expression is often tightly regulated by interactions between transcription factors (TFs) and their DNA cis targets. Yeast one-hybrid (Y1H) is one of the most extensively used methods to discover these interactions. We developed a high-throughput meiosis-directed yeast one-hybrid system using the Magic Markers of the synthetic genetic array analysis. The system has a transcription factor–DNA interaction discovery rate twice as high as the conventional diploid-mating approach and a processing time nearly one-tenth of the haploid-transformation method. The system also offers the highest accuracy in identifying TF–DNA interactions that can be authenticated in vivo by chromatin immunoprecipitation. With these unique features, this meiosis-directed Y1H system is particularly suited for constructing novel and comprehensive genome-scale gene regulatory networks for various organisms.
Footnotes
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[Supplemental material is available for this article.]
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Article published online before print. Article, supplemental material, and publication date are at http://www.genome.org/cgi/doi/10.1101/gr.245951.118.
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Freely available online through the Genome Research Open Access option.
- Received November 2, 2018.
- Accepted June 6, 2019.
This article, published in Genome Research, is available under a Creative Commons License (Attribution-NonCommercial 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/.
