Simple and efficient transgenesis with I-SceI meganuclease in the newt, Cynops pyrrhogaster.
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.
Casco-Robles
Martin Miguel
MM
Graduate School of Life and Environmental Sciences, University of Tsukuba, Tsukuba, Japan.
Yamada
Shouta
S
Miura
Tomoya
T
Chiba
Chikafumi
C
D029845:Amphibian Proteins / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D030801:Animals, Genetically Modified; D004625:Embryo, Nonmammalian / Q000378:metabolism*; D012455:Salamandridae
D029845:Amphibian Proteins