Haralampiev Ivan, Schade Matthias, Chamiolo Jasmine, Jolmes Fabian, Prisner Simon, Witkowski Peter T, Behrent Marie, Hövelmann Felix, Wolff Thorsten, Seitz Oliver, Herrmann Andreas
Institut für Biologie, Molekulare Biophysik, IRI Life Sciences, Humboldt-Universität zu Berlin, Invalidenstrasse 42, 10115, Berlin, Germany.
Institut für Chemie, Bioorganische Synthese, Humboldt-Universität zu Berlin, Brook-Taylor-Strasse 2, 12489, Berlin, Germany.
Chembiochem. 2017 Aug 17;18(16):1589-1592. doi: 10.1002/cbic.201700271. Epub 2017 Jun 27.
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.
甲型流感病毒（IAV）基因组被分割成八个病毒核糖核蛋白，每个核糖核蛋白都表达一条负向的病毒RNA（vRNA）。在感染周期中，高度丰富的单链小病毒RNA（svRNA）以片段特异性方式转录。svRNA的序列和vRNA 5'端的序列相同，并且在所有IAV毒株中高度保守。在此，我们证明这些序列可作为IAV感染的泛选择性传感器的靶标。为此，我们使用了一种带有单个锁核酸取代的互补荧光强制嵌入RNA（IAV QB-FIT）探针来提高亮度。我们通过荧光原位杂交（FISH）证明，该探针适用于且易于用于检测多种禽、猪和人IAV毒株对不同细胞类型的感染，但不适用于其他流感病毒类型。IAV QB-FIT还为表征宿主细胞的不同感染状态提供了一个有用的工具。
