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

Genome Res. 2019 Aug;29(8):1343-1351. doi: 10.1101/gr.245951.118. Epub 2019 Jun 11.

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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • DNA / genetics*
  • DNA / metabolism
  • Gene Expression Regulation
  • Gene Regulatory Networks
  • Genetic Markers
  • Humans
  • Meiosis
  • Microarray Analysis / instrumentation
  • Microarray Analysis / methods*
  • Plasmids / chemistry
  • Plasmids / metabolism
  • Ploidies
  • Populus / cytology
  • Protein Binding
  • Protoplasts / cytology
  • Protoplasts / metabolism
  • Saccharomyces cerevisiae / genetics*
  • Saccharomyces cerevisiae / metabolism
  • Time Factors
  • Transcription Factors / genetics*
  • Transcription Factors / metabolism
  • Two-Hybrid System Techniques*

Substances

  • Genetic Markers
  • Transcription Factors
  • DNA