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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.
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.