Abstract
Base editors (BEs) composed of a cytidine deaminase fused to CRISPR-Cas9 convert cytidine to uridine, leading to single-base-pair substitutions in eukaryotic cells. We delivered BE mRNA or ribonucleoproteins targeting the Dmd or Tyr gene via electroporation or microinjection into mouse zygotes. F0 mice showed nonsense mutations with an efficiency of 44-57% and allelic frequencies of up to 100%, demonstrating an efficient method to generate mice with targeted point mutations.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Base Pairing / genetics
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CRISPR-Associated Proteins / genetics*
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Clustered Regularly Interspaced Short Palindromic Repeats / genetics*
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Cytidine Deaminase / genetics
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Embryo, Mammalian / embryology
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Embryo, Mammalian / physiology*
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Gene Editing / methods*
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Mice
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Mice, Inbred C57BL
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Mice, Inbred ICR
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Mutagenesis, Site-Directed / methods*
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Point Mutation / genetics
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RNA / genetics*
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Recombinant Fusion Proteins / genetics
Substances
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CRISPR-Associated Proteins
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Recombinant Fusion Proteins
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RNA
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Cytidine Deaminase