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Genesis December 1, 2013; 51 (12): 835-43.

Simple and efficient CRISPR/Cas9-mediated targeted mutagenesis in Xenopus tropicalis.

Nakayama T , Fish MB , Fisher M , Oomen-Hajagos J , Thomsen GH , Grainger RM .

We have assessed the efficacy of the recently developed CRISPR/Cas (clustered regularly interspaced short palindromic repeats/CRISPR-associated) system for genome modification in the amphibian Xenopus tropicalis. As a model experiment, targeted mutations of the tyrosinase gene were verified, showing the expected albinism phenotype in injected embryos. We further tested this technology by interrupting the six3 gene, which is required for proper eye and brain formation. Expected eye and brain phenotypes were observed when inducing mutations in the six3 coding regions, as well as when deleting the gene promoter by dual targeting. We describe here a standardized protocol for genome editing using this system. This simple and fast method to edit the genome provides a powerful new reverse genetics tool for Xenopus researchers.

PubMed ID: 24123613
PMC ID: PMC3947545
Article link: Genesis
Grant support: EY018000 NEI NIH HHS , EY022954 NEI NIH HHS , HD064908 NICHD NIH HHS , OD010997 NIH HHS , P40 OD010997 NIH HHS , R01 EY018000 NEI NIH HHS , R01 EY022954 NEI NIH HHS , R03 HD064908 NICHD NIH HHS , T32 GM007964 NIGMS NIH HHS , T32GM007964 NIGMS NIH HHS

Genes referenced: dsp pam rpe six3 sp6 tbx2

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Abu-Daya, 2012, Pubmed, Xenbase [+]

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