COMBINING PROTEIN AND mRNA QUANTIFICATION TO DECIPHER TRANSCRIPTIONAL REGULATION

We combine immunofluorescence and single-molecule fluorescence in situ hybridization (smFISH), followed by automated image analysis, to quantify the concentration of nuclear transcription factors, number of transcription factors bound, and number of nascent mRNAs synthesized at individual gene loci....

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Detalles Bibliográficos
Autores principales: Xu, Heng, Sepúlveda, Leonardo A., Figard, Lauren, Sokac, Anna Marie, Golding, Ido
Formato: Online Artículo Texto
Lenguaje:English
Publicado: 2015
Materias:
Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4521975/
https://www.ncbi.nlm.nih.gov/pubmed/26098021
http://dx.doi.org/10.1038/nmeth.3446
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author Xu, Heng
Sepúlveda, Leonardo A.
Figard, Lauren
Sokac, Anna Marie
Golding, Ido
author_facet Xu, Heng
Sepúlveda, Leonardo A.
Figard, Lauren
Sokac, Anna Marie
Golding, Ido
author_sort Xu, Heng
collection PubMed
description We combine immunofluorescence and single-molecule fluorescence in situ hybridization (smFISH), followed by automated image analysis, to quantify the concentration of nuclear transcription factors, number of transcription factors bound, and number of nascent mRNAs synthesized at individual gene loci. A theoretical model is used to decipher how transcription-factor binding modulates the stochastic kinetics of mRNA production. We demonstrate this approach by examining the regulation of hunchback in the early Drosophila embryo.
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spelling pubmed-45219752016-01-31 COMBINING PROTEIN AND mRNA QUANTIFICATION TO DECIPHER TRANSCRIPTIONAL REGULATION Xu, Heng Sepúlveda, Leonardo A. Figard, Lauren Sokac, Anna Marie Golding, Ido Nat Methods Article We combine immunofluorescence and single-molecule fluorescence in situ hybridization (smFISH), followed by automated image analysis, to quantify the concentration of nuclear transcription factors, number of transcription factors bound, and number of nascent mRNAs synthesized at individual gene loci. A theoretical model is used to decipher how transcription-factor binding modulates the stochastic kinetics of mRNA production. We demonstrate this approach by examining the regulation of hunchback in the early Drosophila embryo. 2015-06-22 2015-08 /pmc/articles/PMC4521975/ /pubmed/26098021 http://dx.doi.org/10.1038/nmeth.3446 Text en http://www.nature.com/authors/editorial_policies/license.html#terms Users may view, print, copy, and download 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
Xu, Heng
Sepúlveda, Leonardo A.
Figard, Lauren
Sokac, Anna Marie
Golding, Ido
COMBINING PROTEIN AND mRNA QUANTIFICATION TO DECIPHER TRANSCRIPTIONAL REGULATION
title COMBINING PROTEIN AND mRNA QUANTIFICATION TO DECIPHER TRANSCRIPTIONAL REGULATION
title_full COMBINING PROTEIN AND mRNA QUANTIFICATION TO DECIPHER TRANSCRIPTIONAL REGULATION
title_fullStr COMBINING PROTEIN AND mRNA QUANTIFICATION TO DECIPHER TRANSCRIPTIONAL REGULATION
title_full_unstemmed COMBINING PROTEIN AND mRNA QUANTIFICATION TO DECIPHER TRANSCRIPTIONAL REGULATION
title_short COMBINING PROTEIN AND mRNA QUANTIFICATION TO DECIPHER TRANSCRIPTIONAL REGULATION
title_sort combining protein and mrna quantification to decipher transcriptional regulation
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4521975/
https://www.ncbi.nlm.nih.gov/pubmed/26098021
http://dx.doi.org/10.1038/nmeth.3446
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