Find in Library

Summary: We have established a new transgenesis protocol based on CRISPR-Cas9, “New and Easy Xenopus Transgenesis (NEXTrans),” and identified a novel safe harbor site in African clawed frogs, Xenopus laevis. We describe steps in detail for the construction of NEXTrans plasmid and guide RNA, CRISPR-Cas9-mediated NEXTrans plasmid integration into the locus, and its validation by genomic PCR. This improved strategy allows us to simply generate transgenic animals that stably express the transgene.For complete details on the use and execution of this protocol, please refer to Shibata et al. (2022).1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.