Abstract
Expression of transgenes within a single generation by direct DNA injection into vertebrate embryos has been plagued by inefficient and nonuniform gene expression. We report a novel strategy for efficient and stable expression of transgenes driven by both ubiquitous and tissue-specific promoters by direct DNA injection into developing Xenopus laevis embryos. This strategy involves flanking expression cassettes of interest with inverted terminal repeat sequences (ITRs) from adeno-associated virus. Our results suggest that the ITR strategy may be generally applicable to other systems, such as zebra fish and embryonic stem cells, and may enable tissue-specific expression of transgenes in problematic contexts.
Publication types
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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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Actins / genetics
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Animals
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Animals, Genetically Modified
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Dependovirus / genetics*
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Embryo, Nonmammalian
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Gene Expression Regulation, Developmental*
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Genetic Engineering / methods*
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Genetic Vectors / genetics
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Green Fluorescent Proteins
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Heart / embryology
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Heart / physiology
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Luminescent Proteins / genetics
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Luminescent Proteins / metabolism
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Organ Specificity
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Plasmids / genetics
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Promoter Regions, Genetic
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Repetitive Sequences, Nucleic Acid
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Somites
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Transgenes*
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Xenopus laevis / embryology*
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beta-Galactosidase / genetics
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beta-Galactosidase / metabolism
Substances
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Actins
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Luminescent Proteins
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Green Fluorescent Proteins
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beta-Galactosidase