Transposon transgenesis in Xenopus.
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.
Yergeau
Donald A
DA
Department of Pathology, St. Jude Children's Research Hospital, Memphis, TN 38105, USA.
Kelley
Clair M
CM
Zhu
Haiqing
H
Kuliyev
Emin
E
Mead
Paul E
PE
D000818:Animals; D030801:Animals, Genetically Modified; D001435:Bacteriophages / Q000378:metabolism; D003433:Crosses, Genetic; D004247:DNA / Q000378:metabolism; D004251:DNA Transposable Elements; D020868:Gene Silencing; D009865:Oocytes / Q000378:metabolism; D010957:Plasmids / Q000378:metabolism; D016133:Polymerase Chain Reaction; D019076:Transgenes; D019895:Transposases / Q000378:metabolism; D014981:Xenopus / Q000378:metabolism*
D004251:DNA Transposable Elements; D004247:DNA; D019895:Transposases