Enhanced yeast one-hybrid assays for high-throughput gene-centered regulatory network mapping.
A major challenge in systems biology is to understand the gene regulatory networks that drive development, physiology and pathology. Interactions between transcription factors and regulatory genomic regions provide the first level of gene control. Gateway-compatible yeast one-hybrid (Y1H) assays present a convenient method to identify and characterize the repertoire of transcription factors that can bind a DNA sequence of interest. To delineate genome-scale regulatory networks, however, large sets of DNA fragments need to be processed at high throughput and high coverage. Here we present enhanced Y1H (eY1H) assays that use a robotic mating platform with a set of improved Y1H reagents and automated readout quantification. We demonstrate that eY1H assays provide excellent coverage and identify interacting transcription factors for multiple DNA fragments in a short time. eY1H assays will be an important tool for mapping gene regulatory networks in Caenorhabditis elegans and other model organisms as well as in humans.
Program in Systems Biology, University of Massachusetts Medical School, Worcester, Massachusetts, USA. john.reece-hoyes@umassmed.edu
John S
JS
Reece-Hoyes
Alos
A
Diallo
Bryan
B
Lajoie
Amanda
A
Kent
Shaleen
S
Shrestha
Sreenath
S
Kadreppa
Colin
C
Pesyna
Job
J
Dekker
Chad L
CL
Myers
Albertha J M
AJ
Walhout
D000818:Animals; D017173:Caenorhabditis elegans / Q000235:genetics / Q000378:metabolism; D004247:DNA / Q000235:genetics; D005786:Gene Expression Regulation; D053263:Gene Regulatory Networks; D057166:High-Throughput Screening Assays; D006801:Humans; D015203:Reproducibility of Results; D049490:Systems Biology; D014157:Transcription Factors / Q000378:metabolism; D020798:Two-Hybrid System Techniques
D014157:Transcription Factors; D004247:DNA