RNA Polymerase I-mediated expression of viral RNA for the rescue of infectious virulent and avirulent Rift Valley fever viruses

A Billecocoq, N Gauliard, N Le May, Richard Michael Elliott, R Flick, M Bouloy

Research output: Contribution to journalArticlepeer-review

95 Citations (Scopus)

Abstract

Rift Valley fever Virus (RVFV, Bunyaviridae, Phlebovirtis) is a mosquito-transmitted arbovirus that causes human and animal diseases in sub-Saharan Africa and was introduced into tire Arabian Peninsula ill 2000. Here, we describe a method of reverse genetics to recover infectious RVFV from transfected plasmids based Oil the use Of the cellular RNA polymerase I promoter to synthesize viral transcripts. We compared its efficiency with a system using T7 RNA polymerase and found that both are equally efficient for the rescue of RVFV generating titers of approx. 10(7) to 10(8) pfu/ml. We used the RNA polymerase 1-based system to rescue both attenuated MP12 and virulent ZH548 strains as well as chimeric MP12-ZH548 viruses, and in addition RVFV expressing reporter proteins. (C) 2008 Elsevier Inc. All rights reserved.

Original languageEnglish
Pages (from-to)377-384
Number of pages8
JournalVirology
Volume378
Issue number2
DOIs
Publication statusPublished - 1 Sept 2008

Keywords

  • bunyavirus
  • phlebovirus
  • Rift Valley fever virus
  • nonstructural protein NSs
  • expression of reporter protein
  • reverse genetics
  • RNA polymerase 1
  • REVERSE-GENETICS SYSTEM
  • NONSTRUCTURAL PROTEIN NSS
  • CLONED CDNA
  • FOREIGN GENE
  • POLYMERASE-I
  • LACKING
  • GENOME
  • BUNYAVIRIDAE
  • ATTENUATION
  • TRANSCRIPTION

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