A spleen necrosis virus-based retroviral vector which expresses two genes from a dicistronic mRNA

Virology. 1992 Feb;186(2):669-75. doi: 10.1016/0042-6822(92)90033-l.

Abstract

We have investigated a novel strategy for coexpressing two genes from a retroviral vector. The 5' nontranslated leader region of at least some picornavirus RNAs contains a sequence that can act as an internal ribosome entry site allowing initiation of translation at a downstream AUG codon in a 5' cap-independent manner. To investigate whether such a sequence can function in the context of a retroviral vector, we constructed a spleen necrosis virus-based vector carrying two selectable marker genes separated by the leader region of encephalomyocarditis virus. This vector was genetically stable and efficiently expressed both markers from a single dicistronic transcript. Since the expression of two genes by other strategies in retroviral vectors can often be problematic, these results offer a promising new approach for the design of "double gene" retroviral vectors.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Base Sequence
  • Blotting, Northern
  • Blotting, Southern
  • Cloning, Molecular
  • DNA, Viral
  • Dogs
  • Encephalomyocarditis virus / genetics
  • Genes
  • Genes, Viral*
  • Genetic Vectors*
  • Kanamycin Kinase
  • Molecular Sequence Data
  • Phosphotransferases (Alcohol Group Acceptor)*
  • Phosphotransferases / genetics
  • Precipitin Tests
  • Proviruses / genetics
  • Proviruses / growth & development
  • RNA, Messenger / genetics*
  • RNA, Messenger / metabolism
  • RNA, Viral / genetics*
  • RNA, Viral / metabolism
  • Reticuloendotheliosis virus / genetics*
  • Ribosomes / metabolism
  • Tumor Cells, Cultured

Substances

  • DNA, Viral
  • RNA, Messenger
  • RNA, Viral
  • Phosphotransferases
  • Phosphotransferases (Alcohol Group Acceptor)
  • hygromycin-B kinase
  • Kanamycin Kinase