Abstract
Here we present a protocol for generating transgenic embryos in Xenopus laevis and Xenopus tropicalis. The method includes three steps: (1) The preparation of high-speed egg extracts, which facilitates the replacement of protamines in sperm nuclei with nucleosomes and decondenses the chromatin of sperm nuclei; (2) The isolation of sperm nuclei; and (3) The mixing of sperm nuclei, restriction enzyme, and high-speed extract in vitro, following by nuclear transplantation into unfertilized eggs to generate the transgenic embryos. This procedure generates non-mosaic transgenic embryos at high frequency and efficiency.
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Ishibashi, S., Kroll, K.L., Amaya, E. (2012). Generating Transgenic Frog Embryos by Restriction Enzyme Mediated Integration (REMI). In: HOPPLER, S., Vize, P. (eds) Xenopus Protocols. Methods in Molecular Biology, vol 917. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-992-1_11
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DOI: https://doi.org/10.1007/978-1-61779-992-1_11
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Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-61779-991-4
Online ISBN: 978-1-61779-992-1
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