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Sensitive visualization of SARS-CoV-2 RNA with CoronaFISH
AbstractThe 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 viral RNA directly in infected cells are critical to analyze its replication cycle, screen for therapeutic molecules or study infections in human tissue. Here, we report the design, validation and initial application of fluorescencein situhybridization (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 (EM). 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.
- Université Paris Diderot France
- University of Paris-Saclay France
- Inserm France
- Institut Pasteur France
Microsoft Academic Graph classification: Coronavirus disease 2019 (COVID-19) medicine.diagnostic_test Oligonucleotide Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA RNA virus Biology biology.organism_classification Virology Transcription (biology) medicine In patient Fluorescence in situ hybridization
[SDV.SPEE]Life Sciences [q-bio]/Santé publique et épidémiologie
[SDV.SPEE]Life Sciences [q-bio]/Santé publique et épidémiologie
Microsoft Academic Graph classification: Coronavirus disease 2019 (COVID-19) medicine.diagnostic_test Oligonucleotide Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA RNA virus Biology biology.organism_classification Virology Transcription (biology) medicine In patient Fluorescence in situ hybridization
citations This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).8 popularity This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.Top 10% influence This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).Average impulse This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.Top 10% citations This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).8 popularity This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.Top 10% influence This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).Average impulse This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.Top 10% Powered byBIP!