Combining protein and mRNA quantification to decipher transcriptional regulation

Heng Xu, Leonardo A. Sepúlveda, Lauren Figard, Anna Marie Sokac, Ido Golding

Research output: Contribution to journalArticlepeer-review

Abstract

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.

Original languageEnglish (US)
Pages (from-to)739-742
Number of pages4
JournalNature Methods
Volume12
Issue number8
DOIs
StatePublished - Jul 30 2015
Externally publishedYes

ASJC Scopus subject areas

  • Biotechnology
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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