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Nat Protoc
2011 May 01;65:600-8. doi: 10.1038/nprot.2011.334.
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Expressing exogenous genes in newts by transgenesis.
Casco-Robles MM
,
Yamada S
,
Miura T
,
Nakamura K
,
Haynes T
,
Maki N
,
Del Rio-Tsonis K
,
Tsonis PA
,
Chiba C
.
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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.
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