A sensitive mNeonGreen reporter system to measure transcriptional dynamics in Drosophila development

Commun Biol. 2020 Nov 12;3(1):663. doi: 10.1038/s42003-020-01375-5.

Abstract

The gene regulatory network governing anterior-posterior axis formation in Drosophila is a well-established paradigm to study transcription in developmental biology. The rapid temporal dynamics of gene expression during early stages of development, however, are difficult to track with standard techniques. We optimized the bright and fast-maturing fluorescent protein mNeonGreen as a real-time, quantitative reporter of enhancer expression. We derive enhancer activity from the reporter fluorescence dynamics with high spatial and temporal resolution, using a robust reconstruction algorithm. By comparing our results with data obtained with the established MS2-MCP system, we demonstrate the higher detection sensitivity of our reporter. We used the reporter to quantify the activity of variants of a simple synthetic enhancer, and observe increased activity upon reduction of enhancer-promoter distance or addition of binding sites for the pioneer transcription factor Zelda. Our reporter system constitutes a powerful tool to study spatio-temporal gene expression dynamics in live embryos.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Binding Sites / genetics
  • Body Patterning / genetics
  • Drosophila Proteins / genetics
  • Drosophila Proteins / metabolism
  • Drosophila* / genetics
  • Drosophila* / growth & development
  • Drosophila* / metabolism
  • Embryo, Nonmammalian
  • Female
  • Fluorescent Dyes / metabolism*
  • Gene Expression Regulation, Developmental / genetics*
  • Gene Regulatory Networks / genetics
  • Genes, Reporter / genetics
  • Male
  • Promoter Regions, Genetic / genetics
  • Transcription Factors / genetics
  • Transcription Factors / metabolism
  • Transcriptome / genetics*

Substances

  • Drosophila Proteins
  • Fluorescent Dyes
  • Transcription Factors