Abstract
Transgenic animals have been indispensable in elucidating and deciphering mechanisms underlying various biological phenomena. In regeneration, transgenic animals expressing fluorescent protein genes have been crucial for identifying the source cells for regeneration and the mechanism of blastema formation. Animals are usually generated by manipulating their genome using various techniques at/in one cell embryo/fertilized egg stage. Here, we describe the generation of germline transgenic axolotls (Ambystoma mexicanum) using the I-SceI meganuclease and Tol2 transposase.
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References
Kragl M, Knapp D, Nacu E, Khattak S, Maden M, Epperlein HH, Tanaka EM (2009) Cells keep a memory of their tissue origin during axolotl limb regeneration. Nature 460:60–65
McHedlishvili L, Mazurov V, Grassme KS, Goehler K, Robl B, Tazaki A, Roensch K, Duemmler A, Tanaka EM (2012) Reconstitution of the central and peripheral nervous system during salamander tail regeneration. Proc Natl Acad Sci U S A 109:E2258–E2266
Sandoval-Guzman T, Wang H, Khattak S, Schuez M, Roensch K, Nacu E, Tazaki A, Joven A, Tanaka EM, Simon A (2014) Fundamental differences in dedifferentiation and stem cell recruitment during skeletal muscle regeneration in two salamander species. Cell Stem Cell 14:174–187
Khattak S, Schuez M, Richter T, Knapp D, Haigo SL, Sandoval-Guzman T, Hradlikova K, Duemmler A, Kerney R, Tanaka EM (2013) Germline transgenic methods for tracking cells and testing gene function during regeneration in the Axolotl. Stem Cell Reports 1:90–103
McHedlishvili L, Epperlein HH, Telzerow A, Tanaka EM (2007) A clonal analysis of neural progenitors during axolotl spinal cord regeneration reveals evidence for both spatially restricted and multipotent progenitors. Development 134:2083–2093
Chen CH, Durand E, Wang J, Zon LI, Poss KD (2013) zebraflash transgenic lines for in vivo bioluminescence imaging of stem cells and regeneration in adult zebrafish. Development 140(24):4988–4997
Gupta V, Poss KD (2012) Clonally dominant cardiomyocytes direct heart morphogenesis. Nature 484:479–484
Choi WY, Gemberling M, Wang J, Holdway JE, Shen MC, Karlstrom RO, Poss KD (2013) In vivo monitoring of cardiomyocyte proliferation to identify chemical modifiers of heart regeneration. Development 140:660–666
Sobkow L, Epperlein HH, Herklotz S, Straube WL, Tanaka EM (2006) A germline GFP transgenic axolotl and its use to track cell fate: dual origin of the fin mesenchyme during development and the fate of blood cells during regeneration. Dev Biol 290:386–397
Khattak S, Richter T, Tanaka EM (2009) Generation of transgenic axolotls (Ambystoma mexicanum). Cold Spring Harb Protoc 2009(8):pdb prot5264. doi:10.1101/pdb.prot5264
Khattak S, Murawala P, Andreas H, Kappert V, Schuez M, Sandoval-Guzman T, Crawford K, Tanaka EM (2014) Optimized axolotl (Ambystoma mexicanum) husbandry, breeding, metamorphosis, transgenesis and tamoxifen-mediated recombination. Nat Protoc 9:529–540
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Khattak, S., Tanaka, E.M. (2015). Transgenesis in Axolotl (Ambystoma mexicanum). In: Kumar, A., Simon, A. (eds) Salamanders in Regeneration Research. Methods in Molecular Biology, vol 1290. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2495-0_21
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DOI: https://doi.org/10.1007/978-1-4939-2495-0_21
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