Analysis of IAV Replication and Co-infection Dynamics by a Versatile RNA Viral Genome Labeling Method
Kouassi, Nancy Mounogou
Date of Issue2017
Lee Kong Chian School of Medicine
Singapore Centre for Environmental Life Sciences and Engineering
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.
IAV Cell Co-infections
© 2017 The Author(s). This paper was published in Cell Reports and is made available as an electronic reprint (preprint) with permission of The Author(s). The published version is available at: [http://dx.doi.org/10.1016/j.celrep.2017.06.021]. One print or electronic copy may be made for personal use only. Systematic or multiple reproduction, distribution to multiple locations via electronic or other means, duplication of any material in this paper for a fee or for commercial purposes, or modification of the content of the paper is prohibited and is subject to penalties under law.