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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,
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 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,
Transposons for cancer gene discovery: Sleeping Beauty and beyond.
2007,
Pubmed Collier,
Hopping around the tumor genome: transposons for cancer gene discovery.
2005,
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,
Excision of the tol2 transposable element of the medaka fish, Oryzias latipes, in zebrafish, Danio rerio.
1998,
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,
Tol2: a versatile gene transfer vector in vertebrates.
2007,
Pubmed
,
Xenbase Kawakami,
A transposon-mediated gene trap approach identifies developmentally regulated genes in zebrafish.
2004,
Pubmed Keng,
Efficient transposition of Tol2 in the mouse germline.
2009,
Pubmed Kitada,
Transposon-tagged mutagenesis in the rat.
2007,
Pubmed Koga,
Transposable element in fish.
1996,
Pubmed Koga,
Amino acid sequence of a putative transposase protein of the medaka fish transposable element Tol2 deduced from mRNA nucleotide sequences.
1999,
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,
Convergence of a head-field selector Otx2 and Notch signaling: a mechanism for lens specification.
2008,
Pubmed
,
Xenbase Ogino,
Highly efficient transgenesis in Xenopus tropicalis using I-SceI meganuclease.
2006,
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,
Site-specific genomic integration in mammalian cells mediated by phage phiC31 integrase.
2001,
Pubmed Thyagarajan,
Mammalian genomes contain active recombinase recognition sites.
2000,
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,
Remobilization of Tol2 transposons in Xenopus tropicalis.
2010,
Pubmed
,
Xenbase Yergeau,
Transgenesis in Xenopus using the Sleeping Beauty transposon system.
2009,
Pubmed
,
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