期刊
NATURE PROTOCOLS
卷 6, 期 5, 页码 600-608出版社
NATURE PUBLISHING GROUP
DOI: 10.1038/nprot.2011.334
关键词
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资金
- Japan Society for the Promotion of Science (JSPS) [20650060, 21300150]
- NIH [EY10540]
- Grants-in-Aid for Scientific Research [20650060, 21300150] Funding Source: KAKEN
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 (similar to 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 45 months.
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