Abstract
The use of CRISPR/Cas9 to knockout genes in zebrafish has been well established. However, to better model many human diseases that are caused by point mutations, a robust methodology for generating desirable DNA base changes is still needed. Recently, Cas9-linked cytidine deaminases (base editors) evolved as a strategy to introduce single base mutations in model organisms. They have been used to convert cytidine to thymine at specific genomic loci. Here we describe a protocol for using the base editing system in zebrafish and its application to reproduce a single base mutation observed in human Ablepharon-Macrostomia Syndrome.
Copyright © 2018 Elsevier Inc. All rights reserved.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Abnormalities, Multiple / genetics
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Animals
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Base Sequence / genetics
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CRISPR-Cas Systems / genetics*
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Cytidine / metabolism
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Cytidine Deaminase / genetics
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Disease Models, Animal
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Embryo, Nonmammalian
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Eye Abnormalities / genetics
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Female
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Gene Editing / methods*
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Humans
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Macrostomia / genetics
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Male
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Mutagenesis, Site-Directed / methods
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Point Mutation / genetics
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RNA, Guide, CRISPR-Cas Systems
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Thymine / metabolism
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Twist-Related Protein 2 / genetics
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Zebrafish / genetics*
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Zebrafish Proteins / genetics
Substances
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RNA, Guide, CRISPR-Cas Systems
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Twist-Related Protein 2
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Zebrafish Proteins
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Cytidine
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Cytidine Deaminase
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Thymine
Supplementary concepts
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Ablepharon macrostomia syndrome