Double subgenomic alphaviruses expressing multiple fluorescent proteins using a Rhopalosiphum padi virus internal ribosome entry site element

Michael R. Wiley, Lisa O. Roberts, Zach N. Adelman, Kevin M. Myles

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Double subgenomic Sindbis virus (dsSINV) vectors are widely used for the expression of proteins, peptides, and RNA sequences. These recombinant RNA viruses permit high level expression of a heterologous sequence in a wide range of animals, tissues, and cells. However, the alphavirus genome structure and replication strategy is not readily amenable to the expression of more than one heterologous sequence. The Rhopalosiphum padi virus (RhPV) genome contains two internal ribosome entry site (IRES) elements that mediate cap-independent translation of the virus nonstructural and structural proteins. Most IRES elements that have been characterized function only in mammalian cells but previous work has shown that the IRES element present in the 59 untranslated region (UTR) of the RhPV genome functions efficiently in mammalian, insect, and plant systems. To determine if the 59 RhPV IRES element could be used to express more than one heterologous sequence from a dsSINV vector, RhPV 59 IRES sequences were placed between genes for two different fluorescent marker proteins in the dsSINV, TE/392J/mcs. While mammalian and insect cells infected with recombinant viruses containing the RhPV sequences expressed both fluorescent marker proteins, only single marker proteins were routinely observed in cells infected with dsSINV vectors in which the RhPV IRES had been replaced by a luciferase fragment, an antisense RhPV IRES, or no intergenic sequence. Thus, we report development of a versatile tool for the expression of multiple sequences in diverse cell types.

Original languageEnglish (US)
Article numbere13924
JournalPloS one
Volume5
Issue number11
DOIs
StatePublished - Dec 9 2010

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Rhopalosiphum padi virus
Alphavirus
ribosomes
Viruses
Sindbis virus
Sindbis Virus
Proteins
Genome
genome
cells
Insects
Genes
viruses
insects
Untranslated Regions
Viral Structural Proteins
proteins
animal tissues
structural proteins
Intergenic DNA

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

Double subgenomic alphaviruses expressing multiple fluorescent proteins using a Rhopalosiphum padi virus internal ribosome entry site element. / Wiley, Michael R.; Roberts, Lisa O.; Adelman, Zach N.; Myles, Kevin M.

In: PloS one, Vol. 5, No. 11, e13924, 09.12.2010.

Research output: Contribution to journalArticle

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