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ZFIN ID: ZDB-PUB-180801-9
In vivo targeted single-nucleotide editing in zebrafish
Tanaka, S., Yoshioka, S., Nishida, K., Hosokawa, H., Kakizuka, A., Maegawa, S.
Date: 2018
Source: Scientific Reports   8: 11423 (Journal)
Registered Authors: Maegawa, Shingo
Keywords: none
MeSH Terms:
  • Animals
  • Base Sequence
  • Cytidine Deaminase/genetics
  • Cytidine Deaminase/metabolism
  • Cytosine/metabolism
  • Embryo, Nonmammalian/metabolism
  • Gene Editing/methods*
  • Heterozygote
  • Inheritance Patterns/genetics
  • Mutation/genetics
  • Nucleotides/genetics*
  • Phenotype
  • Zebrafish/embryology
  • Zebrafish/genetics*
PubMed: 30061715 Full text @ Sci. Rep.
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ABSTRACT
To date, several genome editing technologies have been developed and are widely utilized in many fields of biology. Most of these technologies, if not all, use nucleases to create DNA double-strand breaks (DSBs), raising the potential risk of cell death and/or oncogenic transformation. The risks hinder their therapeutic applications in humans. Here, we show that in vivo targeted single-nucleotide editing in zebrafish, a vertebrate model organism, can be successfully accomplished with the Target-AID system, which involves deamination of a targeted cytidine to create a nucleotide substitution from cytosine to thymine after replication. Application of the system to two zebrafish genes, chordin (chd) and one-eyed pinhead (oep), successfully introduced premature stop codons (TAG or TAA) in the targeted genomic loci. The modifications were heritable and faithfully produced phenocopies of well-known homozygous mutants of each gene. These results demonstrate for the first time that the Target-AID system can create heritable nucleotide substitutions in vivo in a programmable manner, in vertebrates, namely zebrafish.
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