An Improved Reverse Genetics System to Overcome Cell-Type-Dependent Ebola Virus Genome Plasticity

Yoshimi Tsuda, Thomas Hoenen, Logan Banadyga, Carla Weisend, Stacy M. Ricklefs, Stephen F. Porcella, Hideki Ebihara

Research output: Contribution to journalArticlepeer-review

18 Scopus citations

Abstract

Reverse genetics systems represent a key technique for studying replication and pathogenesis of viruses, including Ebola virus (EBOV). During the rescue of recombinant EBOV from Vero cells, a high frequency of mutations was observed throughout the genomes of rescued viruses, including at the RNA editing site of the glycoprotein gene. The influence that such genomic instability could have on downstream uses of rescued virus may be detrimental, and we therefore sought to improve the rescue system. Here we report an improved EBOV rescue system with higher efficiency and genome stability, using a modified full-length EBOV clone in Huh7 cells. Moreover, by evaluating a variety of cells lines, we revealed that EBOV genome instability is cell-type dependent, a fact that has significant implications for the preparation of standard virus stocks. Thus, our improved rescue system will have an impact on both basic and translational research in the filovirus field.

Original languageEnglish (US)
Pages (from-to)S129-S137
JournalJournal of Infectious Diseases
Volume212
DOIs
StatePublished - Oct 1 2015

Keywords

  • Ebola virus
  • RNA editing site
  • mutation
  • reverse genetics system

ASJC Scopus subject areas

  • Immunology and Allergy
  • Infectious Diseases

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