期刊
NATURE PROTOCOLS
卷 9, 期 3, 页码 529-540出版社
NATURE PUBLISHING GROUP
DOI: 10.1038/nprot.2014.040
关键词
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资金
- VolkswagenStiftung, DFG [TA274/3-1, TA274/3-2, TA274/5-1]
- European Research Council Advanced Investigator Grant
- MPI-CBG
- DFG Center for Regenerative Therapies
The axolotl (Mexican salamander, Ambystoma mexicanum) has become a very useful model organism for studying limb and spinal cord regeneration because of its high regenerative capacity. Here we present a protocol for successfully mating and breeding axolotls in the laboratory throughout the year, for metamorphosing axolotls by a single i.p. injection and for axolotl transgenesis using I-SceI meganuclease and the mini Tol2 transposon system. Tol2-mediated transgenesis provides different features and advantages compared with I-SceI-mediated transgenesis, and it can result in more than 30% of animals expressing the transgene throughout their bodies so that they can be directly used for experimentation. By using Tol2-mediated transgenesis, experiments can be performed within weeks (e.g., 5-6 weeks for obtaining 2-3-cm-long larvae) without the need to establish germline transgenic lines (which take 12-18 months). In addition, we describe here tamoxifen-induced Cre-mediated recombination in transgenic axolotls.
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