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Transposon-mediated integration strategies in Xenopus offer simple and robust methods for the generation of germline transgenic animals. Co-injection of fertilized one-cell embryos with plasmid DNA harboring a transposon transgene and synthetic mRNA encoding the cognate transposase enzyme results in mosaic integration of the transposon at early cleavage stages that are frequently passed through the germline in the adult animal. Micro-injection of fertilized embryos is a routine procedure used by many laboratories that use Xenopus as a developmental model and, as such, the transposon transgenesis method can be performed without additional equipment or specialized methodologies. The methods for injecting Xenopus embryos are well documented in the literature so here we provide a step-by-step guide to other aspects of transposon transgenesis, including screening mosaic founders for germline transmission of the transgene and general husbandry considerations related to management of populations of transgenic frogs.
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,
Using phiC31 integrase to make transgenic Xenopus laevis embryos.
2006,
Pubmed
,
Xenbase
Allen,
Bacteriophage phiC31 integrase mediated transgenesis in Xenopus laevis for protein expression at endogenous levels.
2009,
Pubmed
,
Xenbase
Balciunas,
Trapping fish genes with transposons.
2005,
Pubmed
Carlson,
Transposon mutagenesis of the mouse germline.
2003,
Pubmed
Choo,
Zebrafish transgenic Enhancer TRAP line database (ZETRAP).
2006,
Pubmed
Clark,
Transposon vectors for gene-trap insertional mutagenesis in vertebrates.
2004,
Pubmed
Collier,
Hopping around the tumor genome: transposons for cancer gene discovery.
2005,
Pubmed
Collier,
Transposons for cancer gene discovery: Sleeping Beauty and beyond.
2007,
Pubmed
Davidson,
Efficient gene delivery and gene expression in zebrafish using the Sleeping Beauty transposon.
2003,
Pubmed
Ding,
Efficient transposition of the piggyBac (PB) transposon in mammalian cells and mice.
2005,
Pubmed
Doherty,
A flk-1 promoter/enhancer reporter transgenic Xenopus laevis generated using the Sleeping Beauty transposon system: an in vivo model for vascular studies.
2007,
Pubmed
,
Xenbase
Dupuy,
Mammalian mutagenesis using a highly mobile somatic Sleeping Beauty transposon system.
2005,
Pubmed
Dupuy,
Transposition and gene disruption in the male germline of the mouse.
2001,
Pubmed
Etkin,
Distribution, expression and germ line transmission of exogenous DNA sequences following microinjection into Xenopus laevis eggs.
1987,
Pubmed
,
Xenbase
Fraser,
Precise excision of TTAA-specific lepidopteran transposons piggyBac (IFP2) and tagalong (TFP3) from the baculovirus genome in cell lines from two species of Lepidoptera.
1996,
Pubmed
Grabher,
Highly efficient zebrafish transgenesis mediated by the meganuclease I-SceI.
2004,
Pubmed
Grossman,
The piggyBac element is capable of precise excision and transposition in cells and embryos of the mosquito, Anopheles gambiae.
2000,
Pubmed
Groth,
A phage integrase directs efficient site-specific integration in human cells.
2000,
Pubmed
Hackett,
Sleeping beauty transposon-mediated gene therapy for prolonged expression.
2005,
Pubmed
Hamlet,
Tol2 transposon-mediated transgenesis in Xenopus tropicalis.
2006,
Pubmed
,
Xenbase
Horn,
piggyBac-based insertional mutagenesis and enhancer detection as a tool for functional insect genomics.
2003,
Pubmed
Ishibashi,
A method for generating transgenic frog embryos.
2008,
Pubmed
,
Xenbase
Ivics,
Transposons for gene therapy!
2006,
Pubmed
Ivics,
Molecular reconstruction of Sleeping Beauty, a Tc1-like transposon from fish, and its transposition in human cells.
1997,
Pubmed
Kawakami,
Transposon tools and methods in zebrafish.
2005,
Pubmed
Kawakami,
Identification of the Tol2 transposase of the medaka fish Oryzias latipes that catalyzes excision of a nonautonomous Tol2 element in zebrafish Danio rerio.
1999,
Pubmed
Kawakami,
Excision of the tol2 transposable element of the medaka fish, Oryzias latipes, in zebrafish, Danio rerio.
1998,
Pubmed
Kawakami,
A transposon-mediated gene trap approach identifies developmentally regulated genes in zebrafish.
2004,
Pubmed
Kawakami,
Tol2: a versatile gene transfer vector in vertebrates.
2007,
Pubmed
,
Xenbase
Keng,
Efficient transposition of Tol2 in the mouse germline.
2009,
Pubmed
Kitada,
Transposon-tagged mutagenesis in the rat.
2007,
Pubmed
Koga,
Amino acid sequence of a putative transposase protein of the medaka fish transposable element Tol2 deduced from mRNA nucleotide sequences.
1999,
Pubmed
Koga,
Transposable element in fish.
1996,
Pubmed
Kondrychyn,
Genome-wide analysis of Tol2 transposon reintegration in zebrafish.
2009,
Pubmed
Korzh,
Transposons as tools for enhancer trap screens in vertebrates.
2007,
Pubmed
Kotani,
Transposon-mediated gene trapping in zebrafish.
2006,
Pubmed
Kroll,
Transgenic Xenopus embryos from sperm nuclear transplantations reveal FGF signaling requirements during gastrulation.
1996,
Pubmed
,
Xenbase
Largaespada,
Transposon mutagenesis in mice.
2009,
Pubmed
Miskey,
The Frog Prince: a reconstructed transposon from Rana pipiens with high transpositional activity in vertebrate cells.
2003,
Pubmed
,
Xenbase
Nagayoshi,
Insertional mutagenesis by the Tol2 transposon-mediated enhancer trap approach generated mutations in two developmental genes: tcf7 and synembryn-like.
2008,
Pubmed
Ogino,
Highly efficient transgenesis in Xenopus tropicalis using I-SceI meganuclease.
2006,
Pubmed
,
Xenbase
Ogino,
Convergence of a head-field selector Otx2 and Notch signaling: a mechanism for lens specification.
2008,
Pubmed
,
Xenbase
Ogino,
High-throughput transgenesis in Xenopus using I-SceI meganuclease.
2006,
Pubmed
,
Xenbase
Shibano,
Recombinant Tol2 transposase with activity in Xenopus embryos.
2007,
Pubmed
,
Xenbase
Sinzelle,
Generation of trangenic Xenopus laevis using the Sleeping Beauty transposon system.
2006,
Pubmed
,
Xenbase
Smith,
Xenopus laevis transgenesis by sperm nuclear injection.
2006,
Pubmed
,
Xenbase
Smith,
The complete genome sequence of the Streptomyces temperate phage straight phiC31: evolutionary relationships to other viruses.
1999,
Pubmed
Smith,
Isoflurane anesthesia in the African clawed frog (Xenopus laevis).
2000,
Pubmed
,
Xenbase
Sparrow,
A simplified method of generating transgenic Xenopus.
2000,
Pubmed
,
Xenbase
Takeda,
Insertional mutagenesis of the mouse germline with Sleeping Beauty transposition.
2008,
Pubmed
Thyagarajan,
Mammalian genomes contain active recombinase recognition sites.
2000,
Pubmed
Thyagarajan,
Site-specific genomic integration in mammalian cells mediated by phage phiC31 integrase.
2001,
Pubmed
Urasaki,
Efficient transposition of the Tol2 transposable element from a single-copy donor in zebrafish.
2008,
Pubmed
Vignali,
HNF1(beta) is required for mesoderm induction in the Xenopus embryo.
2000,
Pubmed
,
Xenbase
Waldner,
Marking transgenic Xenopus froglets with passive micro transponders.
2007,
Pubmed
,
Xenbase
Woltjen,
piggyBac transposition reprograms fibroblasts to induced pluripotent stem cells.
2009,
Pubmed
Wu,
piggyBac is a flexible and highly active transposon as compared to sleeping beauty, Tol2, and Mos1 in mammalian cells.
2006,
Pubmed
Yergeau,
Injection-mediated transposon transgenesis in Xenopus tropicalis and the identification of integration sites by modified extension primer tag selection (EPTS) linker-mediated PCR.
2007,
Pubmed
,
Xenbase
Yergeau,
Transgenesis in Xenopus using the Sleeping Beauty transposon system.
2009,
Pubmed
,
Xenbase
Yergeau,
Remobilization of Tol2 transposons in Xenopus tropicalis.
2010,
Pubmed
,
Xenbase
