Engineering the Japanese encephalitis virus RNA genome for the expression of foreign genes of various sizes: Implications for packaging capacity and RNA replication efficiency

Sang Im Yun, Yu Jeong Choi, Xiao Fang Yu, Jae Young Song, Young Hak Shin, Young Ran Ju, Seok Yong Kim, Young Min Lee

Research output: Contribution to journalArticle

Abstract

Using the RNA replication machinery of Japanese encephalitis virus (JEV), the authors have established and characterized three strategies for the expression of foreign genes. Initially, ≈11 kb genomic RNA was engineered to express heterologous genes of various sizes by preferentially inserting a new cistron at the beginning of the 3′ nontranslated variable region. RNA transfection yielded recombinant viruses that initiated foreign gene expression after infecting permissive cells. JEV was capable of packaging recombinant genomes as large as ≈15 kb. However, larger genome size was inversely correlated with RNA replication efficiency and cytopathogenicity, with no significant change in infectivity. Second, a variety of self-replicating propagation-deficient viral replicons were constructed by introducing one to three in-frame deletions into the ectodomains of all the structural proteins of JEV. These replicons displayed a spectrum of RNA replication efficiency upon transfection, suggesting that remnant transmembrane domains play a suppressive role in this process. Third, the authors generated a panel of stable packaging cell lines (PCLs) providing all three JEV structural proteins in trans. These PCLs efficiently packaged viral replicon RNAs into single-round infectious viral replicon particles. These JEV-based virus/vector systems may provide useful tools for a variety of biological applications, including foreign gene expression, antiviral compound screening, and genetic immunization.

Original languageEnglish (US)
Pages (from-to)522-535
Number of pages14
JournalJournal of neurovirology
Volume13
Issue number6
DOIs
StatePublished - Nov 1 2007

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Keywords

  • Encapsidation
  • Foreign gene expression
  • Japanese encephalitis virus
  • Packaging cell lines
  • Trans-complementation
  • Viral replicon particles

ASJC Scopus subject areas

  • Neurology
  • Clinical Neurology
  • Cellular and Molecular Neuroscience
  • Virology

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