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Sensitive visualization of SARS-CoV-2 RNA with CoronaFISH
- Source :
- Life Science Alliance, Life Science Alliance, 2022, 5 (4), pp.e202101124. ⟨10.26508/lsa.202101124⟩, Life Science Alliance, Life Science Alliance LLC, 2022, 5 (4), pp.e202101124. ⟨10.26508/lsa.202101124⟩
- Publication Year :
- 2022
- Publisher :
- Life Science Alliance, LLC, 2022.
-
Abstract
- We design an RNA-FISH method to target the plus and minus strands of SARS-CoV-2 RNA, and apply it to image the virus in cell lines, lung tissue, and nasal swabs, by fluorescence and electron microscopy.<br />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.
- Subjects :
- [SDV]Life Sciences [q-bio]
viruses
Health, Toxicology and Mutagenesis
Plant Science
Virus Replication
Antiviral Agents
Sensitivity and Specificity
Biochemistry, Genetics and Molecular Biology (miscellaneous)
03 medical and health sciences
0302 clinical medicine
Cell Line, Tumor
Chlorocebus aethiops
Animals
Humans
Vero Cells
In Situ Hybridization
In Situ Hybridization, Fluorescence
Virus Release
Research Articles
030304 developmental biology
0303 health sciences
Alanine
Ecology
SARS-CoV-2
COVID-19
Reproducibility of Results
respiratory system
Adenosine Monophosphate
respiratory tract diseases
COVID-19 Drug Treatment
3. Good health
[SDV] Life Sciences [q-bio]
Microscopy, Electron
RNA, Viral
Caco-2 Cells
030217 neurology & neurosurgery
Research Article
Subjects
Details
- ISSN :
- 25751077
- Volume :
- 5
- Database :
- OpenAIRE
- Journal :
- Life Science Alliance
- Accession number :
- edsair.doi.dedup.....9964aed5cbfcaa6332af3c93b000effe