Viral sequences enable efficient and tissue-specific expression of transgenes in Xenopus.
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.
Fu
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Y
Department of Medicine, University of California, San Diego, La Jolla 92093-0613, USA.
Wang
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Y
Evans
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SM
D000199:Actins / Q000235:genetics; D000818:Animals; D030801:Animals, Genetically Modified; D000229:Dependovirus / Q000235:genetics*; D004625:Embryo, Nonmammalian; D018507:Gene Expression Regulation, Developmental; D005818:Genetic Engineering / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics; D049452:Green Fluorescent Proteins; D006321:Heart / Q000196:embryology / Q000502:physiology; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D009928:Organ Specificity; D010957:Plasmids / Q000235:genetics; D011401:Promoter Regions, Genetic; D012091:Repetitive Sequences, Nucleic Acid; D019170:Somites; D019076:Transgenes; D014982:Xenopus laevis / Q000196:embryology*; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism
D000199:Actins; D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins; D001616:beta-Galactosidase