Cas9-based genome editing in Xenopus tropicalis.
Xenopus tropicalis has been developed as a model organism for developmental biology, providing a system offering both modern genetics and classical embryology. Recently, the Clustered Regularly Interspaced Short Palindromic Repeats/CRISPR-associated (CRISPR/Cas) system for genome modification has provided an additional tool for Xenopus researchers to achieve simple and efficient targeted mutagenesis. Here, we provide insights into experimental design and procedures permitting successful application of this technique to Xenopus researchers, and offer a general strategy for performing loss-of-function assays in F0 and subsequently F1 embryos.
PubMed ID: 25398349
PMC ID: PMC4284096
Article link: Methods Enzymol
Grant support: EY018000 NEI NIH HHS , EY022954 NEI NIH HHS , HD073179 NICHD NIH HHS , HD080684 NICHD NIH HHS , OD010997 NIH HHS , P40 OD010997 NIH HHS , R01 EY018000 NEI NIH HHS , R01 EY022954 NEI NIH HHS , R01 HD073179 NICHD NIH HHS , R21 HD080684 NICHD NIH HHS