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Sensitive visualization of SARS-CoV-2 RNA with CoronaFISH.
Rensen, Elena; Pietropaoli, Stefano; Mueller, Florian; Weber, Christian; Souquere, Sylvie; Sommer, Sina; Isnard, Pierre; Rabant, Marion; Gibier, Jean-Baptiste; Terzi, Fabiola; Simon-Loriere, Etienne; Rameix-Welti, Marie-Anne; Pierron, Gérard; Barba-Spaeth, Giovanna; Zimmer, Christophe.
  • Rensen E; Institut Pasteur, Université de Paris, CNRS UMR 3691, Imaging and Modeling Unit, Paris, France.
  • Pietropaoli S; Institut Pasteur, Université de Paris, CNRS UMR 3569, Unité de Virologie Structurale, Paris, France.
  • Mueller F; Institut Pasteur, Université de Paris, CNRS UMR 3691, Imaging and Modeling Unit, Paris, France florian.muller@pasteur.fr.
  • Weber C; Institut Pasteur, Université de Paris, CNRS UMR 3691, Imaging and Modeling Unit, Paris, France.
  • Souquere S; Gustave Roussy, AMMICA UMS-3655, Villejuif, France.
  • Sommer S; Institut Pasteur, Université de Paris, CNRS UMR 3569, Unité de Virologie Structurale, Paris, France.
  • Isnard P; Université de Paris, INSERM U1151, CNRS UMR 8253, Institut Necker Enfants Malades, Département "Croissance et Signalisation," Paris, France.
  • Rabant M; Service d'Anatomo-Pathologie, AP-HP, Hôpital Necker Enfants Malades, AP-HP Centre, Paris, France.
  • Gibier JB; Université de Paris, INSERM U1151, CNRS UMR 8253, Institut Necker Enfants Malades, Département "Croissance et Signalisation," Paris, France.
  • Terzi F; Service d'Anatomo-Pathologie, AP-HP, Hôpital Necker Enfants Malades, AP-HP Centre, Paris, France.
  • Simon-Loriere E; Service d'Anatomo-Pathologie, Centre de Biologie Pathologie, CHU Lille, Lille, France.
  • Rameix-Welti MA; Université de Paris, INSERM U1151, CNRS UMR 8253, Institut Necker Enfants Malades, Département "Croissance et Signalisation," Paris, France.
  • Pierron G; Institut Pasteur, Université de Paris, G5 Evolutionary Genomics of RNA Viruses, Paris, France.
  • Barba-Spaeth G; Université Paris-Saclay, INSERM, Université de Versailles St. Quentin, UMR 1173 (2I), Montigny-le-Bretonneux, France.
  • Zimmer C; AP-HP, Université Paris Saclay, Hôpital Ambroise Paré, Laboratoire de Microbiologie, Boulogne-Billancourt, France.
Life Sci Alliance ; 5(4)2022 04.
Article in English | MEDLINE | ID: covidwho-1614505
Preprint
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ABSTRACT
The current COVID-19 pandemic is caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The positive-sense single-stranded RNA virus contains a single linear RNA segment that serves as a template for transcription and replication, leading to the synthesis of positive and negative-stranded viral RNA (vRNA) in infected cells. Tools to visualize vRNA directly in infected cells are critical to analyze the viral replication cycle, screen for therapeutic molecules, or study infections in human tissue. Here, we report the design, validation, and initial application of FISH probes to visualize positive or negative RNA of SARS-CoV-2 (CoronaFISH). We demonstrate sensitive visualization of vRNA in African green monkey and several human cell lines, in patient samples and human tissue. We further demonstrate the adaptation of CoronaFISH probes to electron microscopy. We provide all required oligonucleotide sequences, source code to design the probes, and a detailed protocol. We hope that CoronaFISH will complement existing techniques for research on SARS-CoV-2 biology and COVID-19 pathophysiology, drug screening, and diagnostics.
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Full text: Available Collection: International databases Database: MEDLINE Main subject: Virus Replication / RNA, Viral / In Situ Hybridization, Fluorescence / SARS-CoV-2 / COVID-19 Type of study: Diagnostic study / Clinical Practice Guide Limits: Animals / Humans Language: English Year: 2022 Document Type: Article Affiliation country: Lsa.202101124

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Full text: Available Collection: International databases Database: MEDLINE Main subject: Virus Replication / RNA, Viral / In Situ Hybridization, Fluorescence / SARS-CoV-2 / COVID-19 Type of study: Diagnostic study / Clinical Practice Guide Limits: Animals / Humans Language: English Year: 2022 Document Type: Article Affiliation country: Lsa.202101124