title: Expressing exogenous genes in newts by transgenesis.
abstract: The great regenerative abilities of newts provide the impetus for studies at the molecular level. However, efficient methods for gene regulation have historically been quite limited. Here we describe a protocol for transgenically expressing exogenous genes in the newt Cynops pyrrhogaster. This method is simple: a reaction mixture of I-SceI meganuclease and a plasmid DNA carrying a transgene cassette flanked by the enzyme recognition sites is directly injected into fertilized eggs. The protocol achieves a high efficiency of transgenesis, comparable to protocols used in other animal systems, and it provides a practical number of transgenic newts (∼20% of injected embryos) that survive beyond metamorphosis and that can be applied to regenerative studies. The entire protocol for obtaining transgenic adult newts takes 4-5 months.
authors.affiliation: Graduate School of Life and Environmental Sciences, University of Tsukuba, Ibaraki, 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: Kenta
authors.initials: K
authors.lastname: Nakamura
authors.forename: Tracy
authors.initials: T
authors.lastname: Haynes
authors.forename: Nobuyasu
authors.initials: N
authors.lastname: Maki
authors.forename: Katia
authors.initials: K
authors.lastname: Del Rio-Tsonis
authors.forename: Panagiotis A
authors.initials: PA
authors.lastname: Tsonis
authors.forename: Chikafumi
authors.initials: C
authors.lastname: Chiba
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified / Q000235:genetics; D005260:Female; D018014:Gene Transfer Techniques; D008297:Male; D012455:Salamandridae / Q000235:genetics*