title: A simple method of transgenesis using I-SceI meganuclease in Xenopus.
abstract: Here we present a protocol for generating transgenic embryos in Xenopus using I-SceI meganuclease. This method relies on integration of DNA constructs, containing one or two I-SceI meganuclease sites. It is a simpler method than the REMI method of transgenesis, and it is ideally suited for generating transgenic lines in Xenopus laevis and Xenopus tropicalis. In addition to it being simpler than the REMI method, this protocol also results in single copy integration events rather than tandem concatemers. Although the protocol we describe is for X. tropicalis, the method can also be used to generate transgenic lines in X. laevis. We also describe a convenient method for designing and generating complex constructs for transgenesis, named pTransgenesis, based on the Multisite Gateway(®) cloning, which include I-SceI sites and Tol2 elements to facilitate genome integration.
authors.affiliation: The Healing Foundation Centre, The Faculty of Life Sciences, University of Manchester, Manchester, England, UK.
authors.forename: Shoko
authors.initials: S
authors.lastname: Ishibashi
authors.forename: Nick R
authors.initials: NR
authors.lastname: Love
authors.forename: Enrique
authors.initials: E
authors.lastname: Amaya
mesh_terms: D000818:Animals; D001483:Base Sequence; D006063:Chorionic Gonadotropin / Q000008:administration & dosage; D003001:Cloning, Molecular; D015252:Deoxyribonucleases, Type II Site-Specific / Q000737:chemistry*; D005260:Female; D018014:Gene Transfer Techniques; D006064:Gonadotropins, Equine / Q000008:administration & dosage; D008297:Male; D010957:Plasmids; D012102:Reproductive Control Agents / Q000008:administration & dosage; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry*; D019076:Transgenes; D014981:Xenopus / Q000235:genetics*
chemical_list: D006063:Chorionic Gonadotropin; D006064:Gonadotropins, Equine; D012102:Reproductive Control Agents; D029701:Saccharomyces cerevisiae Proteins; C037725:SCEI protein, S cerevisiae; D015252:Deoxyribonucleases, Type II Site-Specific