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.
CITATION STYLE
Feehan, J. M., Stanar, P., Tam, B. M., Chiu, C., & Moritz, O. L. (2019). Generation and Analysis of Xenopus laevis Models of Retinal Degeneration Using CRISPR/Cas9. In Methods in Molecular Biology (Vol. 1834, pp. 193–207). Humana Press Inc. https://doi.org/10.1007/978-1-4939-8669-9_14
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