A Fluorescent RNA Forced-Intercalation Probe as a Pan-Selective Marker for Influenza A Virus Infection
| Autor: | Peter T. Witkowski, Fabian Jolmes, Matthias Schade, Ivan Haralampiev, Jasmine Chamiolo, Andreas Herrmann, Thorsten Wolff, Felix Hövelmann, Oliver Seitz, Marie Behrent, Simon Prisner |
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| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
viruses Biology 010402 general chemistry medicine.disease_cause Puumala virus 01 natural sciences Biochemistry Genome Virus Madin Darby Canine Kidney Cells 03 medical and health sciences Dogs Orthomyxoviridae Infections medicine Influenza A virus Animals Humans RNA Messenger Locked nucleic acid Molecular Biology In Situ Hybridization Fluorescence Fluorescent Dyes Ribonucleoprotein Base Sequence medicine.diagnostic_test Oligonucleotide Quinolinium Compounds Organic Chemistry RNA RNA Probes Virology Molecular biology Intercalating Agents 0104 chemical sciences 030104 developmental biology Oligodeoxyribonucleotides A549 Cells RNA Viral Molecular Medicine HeLa Cells Fluorescence in situ hybridization |
| Zdroj: | ChemBioChem. 18:1589-1592 |
| ISSN: | 1439-4227 |
| Popis: | The influenza A virus (IAV) genome is segmented into eight viral ribonucleoproteins, each expressing a negatively oriented viral RNA (vRNA). Along the infection cycle, highly abundant single-stranded small viral RNAs (svRNA) are transcribed in a segment-specific manner. The sequences of svRNAs and of the vRNA 5'-ends are identical and highly conserved among all IAV strains. Here, we demonstrate that these sequences can be used as a target for a pan-selective sensor of IAV infection. To this end, we used a complementary fluorescent forced-intercalation RNA (IAV QB-FIT) probe with a single locked nucleic acid substitution to increase brightness. We demonstrated by fluorescence in situ hybridization (FISH) that this probe is suitable and easy to use to detect infection of different cell types by a broad variety of avian, porcine, and human IAV strains, but not by other influenza virus types. IAV QB-FIT also provides a useful tool to characterize different infection states of the host cell. |
| Databáze: | OpenAIRE |
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