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Targeted Genome Sequencing Reveals Varicella-Zoster Virus Open Reading Frame 12 Deletion.
Randall J Cohrs1,2, Katherine S Lee3, Addilynn Beach4
1Department of Neurology, University of Colorado School of Medicine, Aurora, Colorado, USA randall.cohrs@ucdenver.edu.
Journal of Virology
|July 28, 2017
Summary
Varicella-zoster virus (VZV) genetic stability is challenged by laboratory culture. Targeted sequencing revealed a significant deletion in VZV ORF 12, impacting protein expression and Akt phosphorylation.
Area of Science:
- Virology
- Molecular Biology
- Genetics
Background:
- Varicella-zoster virus (VZV) establishes lifelong latent infections.
- Low VZV nucleic acid abundance in neurons hinders latency mechanism studies.
- Optimized targeted capture enriches VZV DNA/cDNA for sequencing.
Purpose of the Study:
- To investigate VZV gene transcription regulation during latency.
- To identify genetic modifications in VZV laboratory strains.
- To assess the impact of VZV ORF 12 deletion on viral protein expression and host cell signaling.
Main Methods:
- Optimized targeted capture protocol for VZV DNA/cDNA enrichment.
- Whole-genome and transcriptome sequencing analysis.
- Analysis of VZV ORF 12 deletion and its consequences on protein expression and Akt phosphorylation.
Main Results:
- A 2,158-bp deletion was detected in VZV ORF 12 of a laboratory strain (VZV32).
- The deletion resulted in abolished ORF 12 and 13 protein expression.
- Akt phosphorylation was inhibited due to the ORF 12 deletion.
Conclusions:
- VZV genome stability is not absolute; laboratory propagation can cause significant genetic alterations.
- Targeted genome sequencing is crucial for authenticating VZV strains cultured in vitro.
- The VZV ORF 12 deletion highlights the need for careful VZV strain characterization in research settings.
