Combining protein and mRNA quantification to decipher transcriptional regulation
Nature methods, 2015•nature.com
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.
(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.
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.
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