Combining protein and mRNA quantification to decipher transcriptional regulation.
Heng XuLeonardo A SepúlvedaLauren FigardAnna Marie SokacIdo GoldingPublished in: Nature methods (2015)
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.
Keyphrases
- transcription factor
- single molecule
- binding protein
- genome wide identification
- dna binding
- genome wide
- atomic force microscopy
- living cells
- machine learning
- high throughput
- copy number
- genome wide analysis
- protein protein
- dna methylation
- genome wide association
- pregnant women
- pregnancy outcomes
- gene expression
- small molecule