Abstract
The 2A-like sequences from members of the picornavirus family were utilized to construct a tricistronic vector bearing the human iduronidase (IDUA) gene along with the firefly luciferase and DsRed2 reporter genes. The 2A-like sequences mediate a cotranslational cleavage event resulting in the release of each individual protein product. Efficient cleavage was observed and all three proteins were functional in vitro and in vivo, allowing for supratherapeutic IDUA enzyme levels and the coexpression of luciferase and DsRed2 expression, which enabled us to track gene expression.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Animals
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Cysteine Endopeptidases / genetics*
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Gene Expression*
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Gene Transfer Techniques
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Genes
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Genes, Reporter*
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Genetic Therapy / methods*
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Genetic Vectors / genetics*
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Iduronidase / metabolism
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In Vitro Techniques
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Luciferases / metabolism
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Luciferases, Firefly
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Luminescent Proteins / genetics
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Mice
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Mice, Inbred NOD
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Mice, SCID
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Models, Genetic
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Mucopolysaccharidoses / metabolism
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NIH 3T3 Cells
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Picornaviridae / genetics*
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Plasmids / metabolism
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Protein Biosynthesis
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Time Factors
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Transfection
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Viral Proteins / genetics*
Substances
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Luminescent Proteins
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Viral Proteins
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fluorescent protein 583
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Luciferases
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Luciferases, Firefly
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Iduronidase
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Cysteine Endopeptidases
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picornain 2A, Picornavirus