A novel synthetic-genetic-array–based yeast one-hybrid system for high discovery rate and short processing time

Author:

Yeh Chung-Shu,Wang Zhifeng,Miao Fang,Ma Hongyan,Kao Chung-Ting,Hsu Tzu-Shu,Yu Jhong-He,Hung Er-Tsi,Lin Chia-Chang,Kuan Chen-Yu,Tsai Ni-Chiao,Zhou Chenguang,Qu Guan-Zheng,Jiang Jing,Liu Guifeng,Wang Jack P.,Li Wei,Chiang Vincent L.,Chang Tien-Hsien,Lin Ying-Chung Jimmy

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.

Funder

National Natural Science Foundation of China

Ministry of Science and Technology

MOST

Academia Sinica Thematic Project

Taiwan MOST

NSFC

U.S. National Science Foundation, Plant Genome Research Program

U.S. Office of Science (Biological and Environmental Research), Department of Energy

Publisher

Cold Spring Harbor Laboratory

Subject

Genetics (clinical),Genetics

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