Mutagenesis of the varicella-zoster virus genome demonstrates that vlt and vlt-orf63 proteins are dispensable for lytic infection

Shirley E. Braspenning, Robert Jan Lebbink, Daniel P. Depledge, Claudia M.E. Schapendonk, Laura A. Anderson, Georges M.G.M. Verjans, Tomohiko Sadaoka, Werner J.D. Ouwendijk

研究成果: ジャーナルへの寄稿学術論文査読

2 被引用数 (Scopus)

抄録

Primary varicella-zoster virus (VZV) infection leads to varicella and the establishment of lifelong latency in sensory ganglion neurons. Reactivation of latent VZV causes herpes zoster, which is frequently associated with chronic pain. Latent viral gene expression is restricted to the VZV latency-associated transcript (VLT) and VLT-ORF63 (VLT63) fusion transcripts. Since VLT and VLT63 encode proteins that are expressed during lytic infection, we investigated whether pVLT and pVLT-ORF63 are essential for VZV replication by performing VZV genome mutagenesis using CRISPR/Cas9 and BAC technologies. We first established that CRISPR/Cas9 can efficiently mutate VZV genomes in lytically VZV-infected cells through targeting non-essential genes ORF8 and ORF11 and subsequently show recovery of viable mutant viruses. By contrast, the VLT region was markedly resistant to CRISPR/Cas9 editing. Whereas most mutants expressed wild-type or N-terminally altered versions of pVLT and pVLT-ORF63, only a minority of the resulting mutant viruses lacked pVLT and pVLT-ORF63 coding potential. Growth curve analysis showed that pVLT/pVLT-ORF63 negative viruses were viable, but impaired in growth in epithelial cells. We confirmed this phenotype independently using BAC-derived pVLT/pVLT-ORF63 negative and repaired viruses. Collectively, these data demonstrate that pVLT and/or pVLT-ORF63 are dispensable for lytic VZV replication but promote efficient VZV infection in epithelial cells.

本文言語英語
論文番号2289
ジャーナルViruses
13
11
DOI
出版ステータス出版済み - 11-2021
外部発表はい

All Science Journal Classification (ASJC) codes

  • 感染症
  • ウイルス学

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