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

Jasmine Chamiolo; Imre Gaspar; Anne Ephrussi; Oliver Seitz

doi:10.1007/978-1-4939-7213-5_18

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.

Authors

Jasmine Chamiolo¹, Imre Gaspar², Anne Ephrussi², Oliver Seitz³

Affiliations

¹ Department of Chemistry, Humboldt University Berlin, Brook-Taylor-Str. 2, D-12489, Berlin, Germany.
² European Molecular Biology Laboratory (EMBL) Heidelberg, Meyerhof Str. 1, 69117, Heidelberg, Germany.
³ Department of Chemistry, Humboldt University Berlin, Brook-Taylor-Str. 2, D-12489, Berlin, Germany. oliver.seitz@chemie.hu-berlin.de.

PMID: 29130204
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