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XB-ART-43033
Proc Natl Acad Sci U S A. April 26, 2011; 108 (17): 7052-7.

Efficient targeted gene disruption in the soma and germ line of the frog Xenopus tropicalis using engineered zinc-finger nucleases.

Young JJ , Cherone JM , Doyon Y , Ankoudinova I , Faraji FM , Lee AH , Ngo C , Guschin DY , Paschon DE , Miller JC , Zhang L , Rebar EJ , Gregory PD , Urnov FD , Harland RM , Zeitler B .


Abstract
The frog Xenopus, an important research organism in cell and developmental biology, currently lacks tools for targeted mutagenesis. Here, we address this problem by genome editing with zinc-finger nucleases (ZFNs). ZFNs directed against an eGFP transgene in Xenopus tropicalis induced mutations consistent with nonhomologous end joining at the target site, resulting in mosaic loss of the fluorescence phenotype at high frequencies. ZFNs directed against the noggin gene produced tadpoles and adult animals carrying up to 47% disrupted alleles, and founder animals yielded progeny carrying insertions and deletions in the noggin gene with no indication of off-target effects. Furthermore, functional tests demonstrated an allelic series of activity between three germ-line mutant alleles. Because ZFNs can be designed against any locus, our data provide a generally applicable protocol for gene disruption in Xenopus.

PubMed ID: 21471457
PMC ID: PMC3084115
Article link: Proc Natl Acad Sci U S A.
Grant support: HD054354 NICHD NIH HHS , R01 HD054354 NICHD NIH HHS , HD054354 NICHD NIH HHS

Genes referenced: nog


References:
Bachiller, 2000, Pubmed[+]


Article Images: [+] show captions

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