Methods Mol Biol 2019 Jan 01;1834:193-207. doi: 10.1007/978-1-4939-8669-9_14.

Generation and Analysis of Xenopus laevis Models of Retinal Degeneration Using CRISPR/Cas9.

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Xenopus laevis have proven to be a useful system for rapid generation and analysis of transgenic models of human retinal disease. However, experimental approaches in this system were limited by lack of a robust knockdown or knockout technology. Here we describe a protocol for generation of Cas9-edited X. laevis embryos. The technique introduces point mutations into the genome of X. laevis resulting in in-frame and out-of-frame insertions and deletions that allow modeling of both dominant and recessive human diseases and efficiently generates gene knockdown and knockout. Our techniques can produce high-frequency gene editing in X. laevis, permitting analysis in the F0 generation.

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