Abstract
Site-specific recombinases are important tools for the modification of mammalian genomes. In conjunction with viral vectors, they can be utilized to mediate site-specific gene insertions in animals and in cell lines which are difficult to transfect. Here we describe a method for the generation and analysis of an adenovirus vector supporting a recombinase-mediated cassette exchange reaction and discuss the advantages and limitations of this approach.
Keywords:
Gene expression; Genome engineering; Genomic target; Site-specific recombinase; TCID50.
MeSH terms
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Caseins / genetics
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Caseins / metabolism
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DNA (Cytosine-5-)-Methyltransferase 1 / genetics
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DNA (Cytosine-5-)-Methyltransferase 1 / metabolism
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DNA Nucleotidyltransferases / genetics
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DNA Nucleotidyltransferases / metabolism
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Dependovirus / genetics*
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Dependovirus / metabolism
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Escherichia coli / genetics
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Escherichia coli / metabolism
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Gene Expression
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Gene Targeting / methods*
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Genes, Reporter
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Genetic Vectors / chemistry
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Genetic Vectors / metabolism
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Green Fluorescent Proteins / genetics
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Green Fluorescent Proteins / metabolism
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HEK293 Cells
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Homologous Recombination*
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Humans
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Integrases / genetics*
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Integrases / metabolism
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Luminescent Proteins / genetics
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Luminescent Proteins / metabolism
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Mutagenesis, Insertional / methods*
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Transfection
Substances
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Caseins
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Luminescent Proteins
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enhanced green fluorescent protein
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fluorescent protein 583
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Green Fluorescent Proteins
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DNA (Cytosine-5-)-Methyltransferase 1
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DNMT1 protein, human
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Cre recombinase
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DNA Nucleotidyltransferases
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Integrases
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Site-specific recombinase