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Methods Mol Biol. 2012;917:129-41. doi: 10.1007/978-1-61779-992-1_7.

Targeted gene disruption with engineered zinc-finger nucleases (ZFNs).

Author information

1
Department of Molecular and Cell Biology, University of California, Berkeley, CA, USA. jyoung528@berkeley.edu

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.

PMID:
22956085
DOI:
10.1007/978-1-61779-992-1_7
[Indexed for MEDLINE]

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