tetano
Editor, Senior Moderator
Cell Rep. 2017 Jul 5;20(1):251-263. doi: 10.1016/j.celrep.2017.06.021.
[h=1]Analysis of IAV Replication and Co-infection Dynamics by a Versatile RNA Viral Genome Labeling Method.[/h] Dou D[SUP]1[/SUP], Hern?ndez-Neuta I[SUP]2[/SUP], Wang H[SUP]1[/SUP], ?stbye H[SUP]1[/SUP], Qian X[SUP]2[/SUP], Thiele S[SUP]3[/SUP], Resa-Infante P[SUP]3[/SUP], Kouassi NM[SUP]3[/SUP], Sender V[SUP]4[/SUP], Hentrich K[SUP]4[/SUP], Mellroth P[SUP]4[/SUP], Henriques-Normark B[SUP]5[/SUP], Gabriel G[SUP]3[/SUP], Nilsson M[SUP]6[/SUP], Daniels R[SUP]7[/SUP].
[h=3]Author information[/h]
[h=3]Abstract[/h] 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.
Copyright ? 2017 The Authors. Published by Elsevier Inc. All rights reserved.
[h=4]KEYWORDS:[/h] 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
PMID: 28683318 DOI: 10.1016/j.celrep.2017.06.021
[h=1]Analysis of IAV Replication and Co-infection Dynamics by a Versatile RNA Viral Genome Labeling Method.[/h] Dou D[SUP]1[/SUP], Hern?ndez-Neuta I[SUP]2[/SUP], Wang H[SUP]1[/SUP], ?stbye H[SUP]1[/SUP], Qian X[SUP]2[/SUP], Thiele S[SUP]3[/SUP], Resa-Infante P[SUP]3[/SUP], Kouassi NM[SUP]3[/SUP], Sender V[SUP]4[/SUP], Hentrich K[SUP]4[/SUP], Mellroth P[SUP]4[/SUP], Henriques-Normark B[SUP]5[/SUP], Gabriel G[SUP]3[/SUP], Nilsson M[SUP]6[/SUP], Daniels R[SUP]7[/SUP].
[h=3]Author information[/h]
[h=3]Abstract[/h] 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.
Copyright ? 2017 The Authors. Published by Elsevier Inc. All rights reserved.
[h=4]KEYWORDS:[/h] 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
PMID: 28683318 DOI: 10.1016/j.celrep.2017.06.021