Simple and efficient transgenesis with I-SceI meganuclease in the newt, Cynops pyrrhogaster

Dev Dyn. 2010 Dec;239(12):3275-84. doi: 10.1002/dvdy.22463.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amphibian Proteins / genetics
  • Amphibian Proteins / metabolism*
  • Animals
  • Animals, Genetically Modified
  • Embryo, Nonmammalian / metabolism*
  • Salamandridae

Substances

  • Amphibian Proteins