Optimized axolotl (Ambystoma mexicanum) husbandry, breeding, metamorphosis, transgenesis and tamoxifen-mediated recombination
Optimized axolotl (Ambystoma mexicanum) husbandry, breeding, metamorphosis, transgenesis and tamoxifen-mediated recombination
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DOI：
10.1038/nprot.2014.040
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发表时间：
2014-03-01
期刊：
影响因子：
14.8
通讯作者：
Tanaka, Elly M.
中科院分区：
文献类型：
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作者：
Khattak, Shahryar;Murawala, Prayag;Tanaka, Elly M.
美西蝾螈（墨西哥蝾螈，Ambystoma mexicanum）已成为一个非常有用的模式生物研究肢体和脊髓再生，因为它的高再生能力。在这里，我们提出了一个协议，成功地交配和繁殖蝾螈在实验室中全年，通过一个单一的IP注射和蝾螈转基因使用I-SceI大范围核酸酶和迷你Tol 2转座子系统的变态蝾螈。与I-SceI介导的转基因相比，Tol 2介导的转基因具有不同的特征和优点，并且它可以导致超过30%的动物在其整个身体中表达转基因，使得它们可以直接用于实验。通过使用Tol 2介导的转基因，可以在数周内进行实验（例如，5-6获得2-3-cm长的幼虫的时间为10 - 20周），而不需要建立生殖系转基因系（需要12-18个月）。此外，我们在这里描述他莫昔芬诱导的Cre介导的重组转基因蝾螈。
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.