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Viruses with RNA Genomes01:29

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RNA viruses are categorized into positive-strand, negative-strand, or double-stranded groups based on their genomic structure and replication mechanisms. This classification dictates how they exploit host cellular machinery for protein synthesis and replication. Some RNA viruses also utilize reverse transcription as part of their life cycle, further diversifying their replication strategies.Positive-Strand RNA VirusesPositive-strand RNA viruses have genomes that function directly as messenger...
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Time-Course Transcriptome Profiling of a Poxvirus Using Long-Read Full-Length Assay.

Dóra Tombácz1,2, István Prazsák1, Gábor Torma1

  • 1Department of Medical Biology, Faculty of Medicine, University of Szeged, 6720 Szeged, Hungary.

Pathogens (Basel, Switzerland)
|August 28, 2021
PubMed
Summary

Long-read sequencing provides complete viral transcriptomes, overcoming limitations of older methods for vaccinia virus (VACV) gene expression analysis. This advanced technique accurately identifies complex transcriptional start and end sites.

Keywords:
gene expressionlong-read sequencingnanopore sequencingtranscriptome profilingvaccinia virus

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Area of Science:

  • Virology
  • Genomics
  • Molecular Biology

Background:

  • First- and second-generation sequencing techniques yield incomplete viral transcriptomes.
  • Short read lengths hinder the distinction of transcript isoforms, polycistronic RNAs, and transcriptional overlaps.
  • These methods are insensitive for identifying transcriptional start sites (TSSs) and end sites (TESs), especially for complex transcripts.

Purpose of the Study:

  • To investigate the time-lapse transcriptome patterns of vaccinia virus (VACV) gene expression.
  • To leverage long-read sequencing for a comprehensive analysis of viral transcriptomes.
  • To address the complexity arising from VACV's diverse TSSs, TESs, and polycistronism.

Main Methods:

  • Application of single-molecule, real-time (SMRT) sequencing.
  • Utilizing nanopore-based sequencing technologies.
  • Analysis of time-lapse VACV gene expression patterns.

Main Results:

  • Long-read sequencing enables the assembly of complete viral transcriptome atlases.
  • VACV transcriptome exhibits high diversity in TSSs and TESs.
  • A high degree of polycistronism contributes to significant transcriptome complexity in VACV.

Conclusions:

  • Long-read sequencing is essential for accurate and complete viral transcriptome analysis.
  • This approach overcomes the limitations of previous sequencing technologies for complex viral genomes.
  • The study provides novel insights into the intricate gene expression dynamics of vaccinia virus.