Reference - Detail
| RRC ID | 77294 |
|---|---|
| Author | Suzuki M, Iida M, Hayashi T, Suzuki KT. |
| Title | CRISPR-Cas9-Based Functional Analysis in Amphibians: Xenopus laevis, Xenopus tropicalis, and Pleurodeles waltl. |
| Journal | Methods Mol Biol |
| Abstract |
Amphibians have made many fundamental contributions to our knowledge, from basic biology to biomedical research on human diseases. Current genome editing tools based on the CRISPR-Cas system enable us to perform gene functional analysis in vivo, even in non-model organisms. We introduce here a highly efficient and easy protocol for gene knockout, which can be used in three different amphibians seamlessly: Xenopus laevis, Xenopus tropicalis, and Pleurodeles waltl. As it utilizes Cas9 ribonucleoprotein complex (RNP) for injection, this cloning-free method enables researchers to obtain founder embryos with a nearly complete knockout phenotype within a week. To evaluate somatic mutation rate and its correlation to the phenotype of a Cas9 RNP-injected embryo (crispant), we also present accurate and cost-effective genotyping methods using pooled amplicon-sequencing and a user-friendly web-based tool. |
| Volume | 2637 |
| Pages | 341-357 |
| Published | 2023-2-12 |
| DOI | 10.1007/978-1-0716-3016-7_26 |
| PMID | 36773159 |
| MeSH | Animals CRISPR-Cas Systems* / genetics Gene Editing / methods Humans Pleurodeles* / genetics Xenopus / genetics Xenopus laevis / genetics |
| Altmetric score |
オルトメトリクス指標項目
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| The most frequently cited source | X(Twitter) |
| Total number of mentions | 3 |
| Altmetric score changes over past 6months | 0.0 |
| Resource | |
| Clawed frogs / Newts | Xenopus tropicalis Pleurodeles waltl |