A spleen necrosis virus-based retroviral vector which expresses two genes from a dicistronic mRNA.
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.
Koo
H M
HM
Department of Molecular Genetics and Microbiology, Robert Wood Johnson Medical School, University of Medicine and Dentistry of New Jersey, Piscataway 08854-5635.
Brown
A M
AM
Kaufman
R J
RJ
Prorock
C M
CM
Ron
Y
Y
Dougherty
J P
JP
D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D015139:Blotting, Southern; D003001:Cloning, Molecular; D004279:DNA, Viral; D004285:Dogs; D004686:Encephalomyocarditis virus / Q000235:genetics; D005796:Genes; D005814:Genes, Viral; D005822:Genetic Vectors; D019868:Kanamycin Kinase; D008969:Molecular Sequence Data; D010770:Phosphotransferases / Q000235:genetics; D017853:Phosphotransferases (Alcohol Group Acceptor); D011233:Precipitin Tests; D011533:Proviruses / Q000235:genetics / Q000254:growth & development; D012333:RNA, Messenger / Q000235:genetics* / Q000378:metabolism; D012367:RNA, Viral / Q000235:genetics* / Q000378:metabolism; D001356:Reticuloendotheliosis virus / Q000235:genetics*; D012270:Ribosomes / Q000378:metabolism; D014407:Tumor Cells, Cultured
D004279:DNA, Viral; D012333:RNA, Messenger; D012367:RNA, Viral; D010770:Phosphotransferases; D017853:Phosphotransferases (Alcohol Group Acceptor); C063242:hygromycin-B kinase; D019868:Kanamycin Kinase