Analysis of IAV Replication and Co-infection Dynamics by a Versatile RNA Viral Genome Labeling Method - PubMed (original) (raw)

. 2017 Jul 5;20(1):251-263.

doi: 10.1016/j.celrep.2017.06.021.

Iván Hernández-Neuta 2, Hao Wang 1, Henrik Östbye 1, Xiaoyan Qian 2, Swantje Thiele 3, Patricia Resa-Infante 3, Nancy Mounogou Kouassi 3, Vicky Sender 4, Karina Hentrich 4, Peter Mellroth 4, Birgitta Henriques-Normark 5, Gülsah Gabriel 3, Mats Nilsson 6, Robert Daniels 7

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Analysis of IAV Replication and Co-infection Dynamics by a Versatile RNA Viral Genome Labeling Method

Dan Dou et al. Cell Rep. 2017.

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Abstract

Genome delivery to the proper cellular compartment for transcription and replication is a primary goal of viruses. However, methods for analyzing viral genome localization and differentiating genomes with high identity are lacking, making it difficult to investigate entry-related processes and co-examine heterogeneous RNA viral populations. Here, we present an RNA labeling approach for single-cell analysis of RNA viral replication and co-infection dynamics in situ, which uses the versatility of padlock probes. We applied this method to identify influenza A virus (IAV) infections in cells and lung tissue with single-nucleotide specificity and to classify entry and replication stages by gene segment localization. Extending the classification strategy to co-infections of IAVs with single-nucleotide variations, we found that the dependence on intracellular trafficking places a time restriction on secondary co-infections necessary for genome reassortment. Altogether, these data demonstrate how RNA viral genome labeling can help dissect entry and co-infections.

Keywords: IAV cell co-infections; IAV entry; IAV replication cycle; RNA labeling; RNA viruses; influenza A virus; single-cell IAV genome trafficking; single-nucleotide specificity; vRNAs; viral genome labeling and localization.

Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.

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