In Vivo Visualization and Function Probing of Transport mRNPs Using Injected FIT Probes

Methods Mol Biol. 2018;1649:273-287. doi: 10.1007/978-1-4939-7213-5_18.

Abstract

Fluorogenic hybridization methods, such as the use of FIT probes, enable the in vivo detection of specific mRNAs transcribed from their endogenous, genetically nonmodified loci. Here, we describe the design, synthesis and injection of nuclease resistant FIT probes into developing Drosophila oocytes to detect endogenous localizing mRNAs as wells as to probe function of structural RNA elements.

Keywords: Forced intercalation of thiazole orange; Injection; Live cell imaging; Locked nucleic acid; mRNA transport.

MeSH terms

  • Animals
  • Benzothiazoles / chemistry*
  • Dissection
  • Drosophila melanogaster / cytology
  • Drosophila melanogaster / metabolism*
  • Female
  • Imaging, Three-Dimensional
  • In Situ Hybridization, Fluorescence / methods*
  • Intercalating Agents / chemistry*
  • Microinjections
  • Oocytes / cytology
  • Oocytes / metabolism
  • Protein Transport
  • Quinolines / chemistry*
  • RNA Probes / metabolism*
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Ribonucleoproteins / metabolism*

Substances

  • Benzothiazoles
  • Intercalating Agents
  • Quinolines
  • RNA Probes
  • RNA, Messenger
  • Ribonucleoproteins
  • messenger ribonucleoprotein
  • thiazole orange