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Enhanced yeast one-hybrid (eY1H) 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 pre...
Autores principales: | , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
2011
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3235803/ https://www.ncbi.nlm.nih.gov/pubmed/22037705 http://dx.doi.org/10.1038/nmeth.1748 |
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author | Reece-Hoyes, John S Diallo, Alos Lajoie, Bryan Kent, Amanda Shrestha, Shaleen Kadreppa, Sreenath Pesyna, Colin Dekker, Job Myers, Chad L Walhout, Albertha J M |
author_facet | Reece-Hoyes, John S Diallo, Alos Lajoie, Bryan Kent, Amanda Shrestha, Shaleen Kadreppa, Sreenath Pesyna, Colin Dekker, Job Myers, Chad L Walhout, Albertha J M |
author_sort | Reece-Hoyes, John S |
collection | PubMed |
description | 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 utilize 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 amount of time. eY1H assays will be an important tool for gene regulatory network mapping in Caenorhabditis elegans and other model organisms, as well as humans. |
format | Online Article Text |
id | pubmed-3235803 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2011 |
record_format | MEDLINE/PubMed |
spelling | pubmed-32358032012-06-01 Enhanced yeast one-hybrid (eY1H) assays for high-throughput gene-centered regulatory network mapping Reece-Hoyes, John S Diallo, Alos Lajoie, Bryan Kent, Amanda Shrestha, Shaleen Kadreppa, Sreenath Pesyna, Colin Dekker, Job Myers, Chad L Walhout, Albertha J M Nat Methods Article 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 utilize 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 amount of time. eY1H assays will be an important tool for gene regulatory network mapping in Caenorhabditis elegans and other model organisms, as well as humans. 2011-10-30 /pmc/articles/PMC3235803/ /pubmed/22037705 http://dx.doi.org/10.1038/nmeth.1748 Text en Users may view, print, copy, download and text and data- mine the content in such documents, for the purposes of academic research, subject always to the full Conditions of use: http://www.nature.com/authors/editorial_policies/license.html#terms |
spellingShingle | Article Reece-Hoyes, John S Diallo, Alos Lajoie, Bryan Kent, Amanda Shrestha, Shaleen Kadreppa, Sreenath Pesyna, Colin Dekker, Job Myers, Chad L Walhout, Albertha J M Enhanced yeast one-hybrid (eY1H) assays for high-throughput gene-centered regulatory network mapping |
title | Enhanced yeast one-hybrid (eY1H) assays for high-throughput gene-centered regulatory network mapping |
title_full | Enhanced yeast one-hybrid (eY1H) assays for high-throughput gene-centered regulatory network mapping |
title_fullStr | Enhanced yeast one-hybrid (eY1H) assays for high-throughput gene-centered regulatory network mapping |
title_full_unstemmed | Enhanced yeast one-hybrid (eY1H) assays for high-throughput gene-centered regulatory network mapping |
title_short | Enhanced yeast one-hybrid (eY1H) assays for high-throughput gene-centered regulatory network mapping |
title_sort | enhanced yeast one-hybrid (ey1h) assays for high-throughput gene-centered regulatory network mapping |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3235803/ https://www.ncbi.nlm.nih.gov/pubmed/22037705 http://dx.doi.org/10.1038/nmeth.1748 |
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