LlamaTags: A Versatile Tool to Image Transcription Factor Dynamics in Live Embryos

Cell. 2018 Jun 14;173(7):1810-1822.e16. doi: 10.1016/j.cell.2018.03.069. Epub 2018 May 10.

Abstract

Embryonic cell fates are defined by transcription factors that are rapidly deployed, yet attempts to visualize these factors in vivo often fail because of slow fluorescent protein maturation. Here, we pioneer a protein tag, LlamaTag, which circumvents this maturation limit by binding mature fluorescent proteins, making it possible to visualize transcription factor concentration dynamics in live embryos. Implementing this approach in the fruit fly Drosophila melanogaster, we discovered stochastic bursts in the concentration of transcription factors that are correlated with bursts in transcription. We further used LlamaTags to show that the concentration of protein in a given nucleus heavily depends on transcription of that gene in neighboring nuclei; we speculate that this inter-nuclear signaling is an important mechanism for coordinating gene expression to delineate straight and sharp boundaries of gene expression. Thus, LlamaTags now make it possible to visualize the flow of information along the central dogma in live embryos.

Keywords: Drosophila melanogaster; Fushi-Tarazu; cell-to-cell communication; even-skipped; fluorescent protein maturation; live imaging; snail; transcription; transcription factors; translation.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Cell Nucleus / metabolism
  • Clustered Regularly Interspaced Short Palindromic Repeats / genetics
  • Drosophila Proteins / genetics*
  • Drosophila Proteins / metabolism
  • Drosophila melanogaster / growth & development
  • Drosophila melanogaster / metabolism*
  • Embryo, Nonmammalian / metabolism
  • Embryo, Nonmammalian / pathology
  • Gene Editing / methods*
  • Gene Expression Regulation, Developmental
  • Green Fluorescent Proteins / genetics
  • Microscopy, Confocal
  • Transcription Factors / genetics*
  • Transcription Factors / metabolism

Substances

  • Drosophila Proteins
  • Transcription Factors
  • Green Fluorescent Proteins