title: Simple and efficient transgenesis with I-SceI meganuclease in the newt, Cynops pyrrhogaster.
abstract: Newts have been recognized as an ideal model for body-parts regeneration after traumatic injury since the 18(th) century. However, molecular mechanisms underlying regeneration remain a mystery because of technical limitations. In the current study, to break this obstacle, we established a simple and efficient transgenic protocol for the newt Cynops pyrrhogaster by adapting an I-SceI microinjection technique, as well as a two-aquarium-tank (TAT) system that allows us to constantly obtain fertilized eggs in the laboratory for transgenesis. Following our protocol, ∼ 20% of injected embryos would exhibit non-mosaic widespread transgene expression and survive beyond metamorphosis. This anticipated success rate is about 10 times higher than that obtained by previous protocols, reaching a practical level. Therefore, our transgenic protocol in conjunction with the TAT-system could provide a key technique to open the way to uncover the long mystery underlying body-parts regeneration of newts.
authors.affiliation: Graduate School of Life and Environmental Sciences, University of Tsukuba, Tsukuba, Japan.
authors.forename: Martin Miguel
authors.initials: MM
authors.lastname: Casco-Robles
authors.forename: Shouta
authors.initials: S
authors.lastname: Yamada
authors.forename: Tomoya
authors.initials: T
authors.lastname: Miura
authors.forename: Chikafumi
authors.initials: C
authors.lastname: Chiba
mesh_terms: D029845:Amphibian Proteins / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D030801:Animals, Genetically Modified; D004625:Embryo, Nonmammalian / Q000378:metabolism*; D012455:Salamandridae
chemical_list: D029845:Amphibian Proteins