Abstract
Reverse genetics in Xenopus has been limited to knockdown strategies using antisense morpholino oligonucleotides (MOs). Recently, engineered zinc-finger nucleases have been used to induce targeted mutations resulting in null alleles. Zinc-finger nuclease (ZFN) technology has been adapted to induce null mutations in many systems previously refractory to targeted gene inactivation. Here we provide a general protocol for inducing targeted mutations in Xenopus tropicalis using ZFNs, a method to detect resulting mutations, and the steps to generate homozygous mutant embryos.
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We wish to thank the Conlon and Moens labs for the celery extract preparation protocols.
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Young, J.J., Harland, R.M. (2012). Targeted Gene Disruption with Engineered Zinc-Finger Nucleases (ZFNs). 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_7
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DOI: https://doi.org/10.1007/978-1-61779-992-1_7
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