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Bacteriophage phiC31 encodes an integrase that can mediate the insertion of extrachromosomal DNA into genomic DNA. Here we show that the coinjection of mRNA encoding phiC31 integrase with plasmid DNA encoding the green fluorescent protein (GFP) can be used to generate transgenic X. laevis embryos. Despite integration into the genome, appropriate promoter expression required modification of the reporter plasmid by bracketing the GFP reporter gene with tandem copies of the chicken beta-globin 5' HS4 insulator to relieve silencing owing to chromatin position effects. These experiments demonstrate that the integration of insulated gene sequences using phiC31 integrase can be used to efficiently create transgenic embryos in X. laevis and may increase the practical use of phiC31 integrase in other systems as well.
Bell,
Stopped at the border: boundaries and insulators.
1999, Pubmed
Bell,
Stopped at the border: boundaries and insulators.
1999,
Pubmed
Belteki,
Site-specific cassette exchange and germline transmission with mouse ES cells expressing phiC31 integrase.
2003,
Pubmed
Chalberg,
phiC31 integrase confers genomic integration and long-term transgene expression in rat retina.
2005,
Pubmed
Chung,
A 5' element of the chicken beta-globin domain serves as an insulator in human erythroid cells and protects against position effect in Drosophila.
1993,
Pubmed
Dagle,
Targeted degradation of mRNA in Xenopus oocytes and embryos directed by modified oligonucleotides: studies of An2 and cyclin in embryogenesis.
1990,
Pubmed
,
Xenbase
Groth,
A phage integrase directs efficient site-specific integration in human cells.
2000,
Pubmed
Groth,
Construction of transgenic Drosophila by using the site-specific integrase from phage phiC31.
2004,
Pubmed
Held,
In vivo correction of murine hereditary tyrosinemia type I by phiC31 integrase-mediated gene delivery.
2005,
Pubmed
Kroll,
Transgenic Xenopus embryos from sperm nuclear transplantations reveal FGF signaling requirements during gastrulation.
1996,
Pubmed
,
Xenbase
Kuhn,
Genomic insulators: connecting properties to mechanism.
2003,
Pubmed
Lutz,
A novel approach to plastid transformation utilizes the phiC31 phage integrase.
2004,
Pubmed
Offield,
The development of Xenopus tropicalis transgenic lines and their use in studying lens developmental timing in living embryos.
2000,
Pubmed
,
Xenbase
Olivares,
Site-specific genomic integration produces therapeutic Factor IX levels in mice.
2002,
Pubmed
Pikaart,
Loss of transcriptional activity of a transgene is accompanied by DNA methylation and histone deacetylation and is prevented by insulators.
1998,
Pubmed
Rebagliati,
Identification and cloning of localized maternal RNAs from Xenopus eggs.
1985,
Pubmed
,
Xenbase
Smith,
The complete genome sequence of the Streptomyces temperate phage straight phiC31: evolutionary relationships to other viruses.
1999,
Pubmed
Smith,
Frog transgenesis made simple.
2005,
Pubmed
,
Xenbase
Sparrow,
A simplified method of generating transgenic Xenopus.
2000,
Pubmed
,
Xenbase
Thomason,
Gene insertion and replacement in Schizosaccharomyces pombe mediated by the Streptomyces bacteriophage phiC31 site-specific recombination system.
2001,
Pubmed
Thorpe,
Control of directionality in the site-specific recombination system of the Streptomyces phage phiC31.
2000,
Pubmed
Thorpe,
In vitro site-specific integration of bacteriophage DNA catalyzed by a recombinase of the resolvase/invertase family.
1998,
Pubmed
Thyagarajan,
Site-specific genomic integration in mammalian cells mediated by phage phiC31 integrase.
2001,
Pubmed
Vize,
Assays for gene function in developing Xenopus embryos.
1991,
Pubmed
,
Xenbase
West,
Insulators: many functions, many mechanisms.
2002,
Pubmed
