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. © 2012 Springer Science+Business Media, LLC.
CITATION STYLE
Young, J. J., & Harland, R. M. (2012). Targeted gene disruption with engineered Zinc-Finger Nucleases (ZFNs). Methods in Molecular Biology, 917, 129–141. https://doi.org/10.1007/978-1-61779-992-1_7
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