Click here to close
Hello! We notice that you are using Internet Explorer, which is not supported by Xenbase and may cause the site to display incorrectly.
We suggest using a current version of Chrome,
FireFox, or Safari.
???displayArticle.abstract???
The two most common methods used to generate transgenic Xenopus embryos, restriction enzyme-mediated insertion, and I-SceI meganuclease take advantage of relatively common but spatially unpredictable double-stranded breaks in sperm, egg, or early embryo genomes. These methods also tend to insert multimeric copies of the transgene. An alternative is to use bacteriophage- or transposon-derived integrase or recombinase to mediate more site-specific insertion of the transgene. The use of phiC31 integrase requires a defined sequence for insertion and is compatible with insertion of a single copy of the transgene. We describe the protocol we use to facilitate phiC31 integrase transgene insertion including the use of insulator sequences to reduce position effect disruption of transgene activity.
Allen,
Transgenic Xenopus laevis embryos can be generated using phiC31 integrase.
2005, Pubmed,
Xenbase
Allen,
Transgenic Xenopus laevis embryos can be generated using phiC31 integrase.
2005,
Pubmed
,
Xenbase
Allen,
Bacteriophage phiC31 integrase mediated transgenesis in Xenopus laevis for protein expression at endogenous levels.
2009,
Pubmed
,
Xenbase
Allen,
Using phiC31 integrase to make transgenic Xenopus laevis embryos.
2006,
Pubmed
,
Xenbase
Bell,
Insulators and boundaries: versatile regulatory elements in the eukaryotic genome.
2001,
Pubmed
Chesneau,
Transgenesis procedures in Xenopus.
2008,
Pubmed
,
Xenbase
Combes,
The streptomyces genome contains multiple pseudo-attB sites for the (phi)C31-encoded site-specific recombination system.
2002,
Pubmed
Groth,
Construction of transgenic Drosophila by using the site-specific integrase from phage phiC31.
2004,
Pubmed
Groth,
A phage integrase directs efficient site-specific integration in human cells.
2000,
Pubmed
Groth,
Phage integrases: biology and applications.
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
Lister,
Transgene excision in zebrafish using the phiC31 integrase.
2010,
Pubmed
Ogino,
Highly efficient transgenesis in Xenopus tropicalis using I-SceI meganuclease.
2006,
Pubmed
,
Xenbase
Olivares,
Site-specific genomic integration produces therapeutic Factor IX levels in mice.
2002,
Pubmed
Ortiz-Urda,
Stable nonviral genetic correction of inherited human skin disease.
2002,
Pubmed
Pan,
I-SceI meganuclease-mediated transgenesis in Xenopus.
2006,
Pubmed
,
Xenbase
Saitoh,
Structural and functional conservation at the boundaries of the chicken beta-globin domain.
2000,
Pubmed
Smith,
Synapsis and DNA cleavage in phiC31 integrase-mediated site-specific recombination.
2004,
Pubmed
Smith,
The complete genome sequence of the Streptomyces temperate phage straight phiC31: evolutionary relationships to other viruses.
1999,
Pubmed
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
Thyagarajan,
Mammalian genomes contain active recombinase recognition sites.
2000,
Pubmed
West,
Insulators: many functions, many mechanisms.
2002,
Pubmed
