Supersensitive Multifluorophore RNA-FISH for Early Virus Detection and Flow-FISH by Using Click Chemistry

Raddaoui N, Croce S, Geiger F, Borodavka A, Möckl L, Stazzoni S, Viverge B, Bräuchle C, Frischmuth T, Engelke H, Carell T (2020)

Publication Type: Journal article

Publication year: 2020

Journal

ChemBioChem Wiley-VCH Verlag

Book Volume: 21

Pages Range: 2214-2218

Journal Issue: 15

DOI: 10.1002/cbic.202000081

Abstract

The reliable detection of transcription events through the quantification of the corresponding mRNA is of paramount importance for the diagnostics of infections and diseases. The quantification and localization analysis of the transcripts of a particular gene allows disease states to be characterized more directly compared to an analysis on the transcriptome wide level. This is particularly needed for the early detection of virus infections as now required for emergent viral diseases, e. g. Covid-19. In situ mRNA analysis, however, is a formidable challenge and currently performed with sets of single-fluorophore-containing oligonucleotide probes that hybridize to the mRNA in question. Often a large number of probe strands (>30) are required to get a reliable signal. The more oligonucleotide probes are used, however, the higher the potential off-target binding effects that create background noise. Here, we used click chemistry and alkyne-modified DNA oligonucleotides to prepare multiple-fluorophore-containing probes. We found that these multiple-dye probes allow reliable detection and direct visualization of mRNA with only a very small number (5–10) of probe strands. The new method enabled the in situ detection of viral transcripts as early as 4 hours after infection.

Authors with CRIS profile

Leonhard Möckl

Involved external institutions

Ludwig-Maximilians-Universität (LMU) DE Germany (DE) University of Leeds GB United Kingdom (GB) LMU Klinikum DE Germany (DE)

How to cite

APA:

Raddaoui, N., Croce, S., Geiger, F., Borodavka, A., Möckl, L., Stazzoni, S.,... Carell, T. (2020). Supersensitive Multifluorophore RNA-FISH for Early Virus Detection and Flow-FISH by Using Click Chemistry. ChemBioChem, 21(15), 2214-2218. https://doi.org/10.1002/cbic.202000081

MLA:

Raddaoui, Nada, et al. "Supersensitive Multifluorophore RNA-FISH for Early Virus Detection and Flow-FISH by Using Click Chemistry." ChemBioChem 21.15 (2020): 2214-2218.

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