1Imagerie et Modélisation - Imaging and Modeling (Département Biologie computationnelle,
Département de Biologie cellulaire et Infection,
25-28, rue du docteur Roux, 75724 Paris Cedex 15 - France)
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.
Elena Rensen, Stefano Pietropaoli, Florian Mueller, Christian Weber, Sylvie Souquere, et al.. Sensitive visualization of SARS-CoV-2 RNA with CoronaFISH. Life Science Alliance, Life Science Alliance LLC, 2022, 5 (4), pp.e202101124. ⟨10.26508/lsa.202101124⟩. ⟨pasteur-03518024v2⟩