ZFIN ID: ZDB-PUB-130124-4
A large-scale in vivo analysis reveals that TALENs are significantly more mutagenic than ZFNs generated using context-dependent assembly
Chen, S., Oikonomou, G., Chiu, C.N., Niles, B.J., Liu, J., Lee, D.A., Antoshechkin, I., and Prober, D.A.
Date: 2013
Source: Nucleic acids research   41(4): 2769-2778 (Journal)
Registered Authors: Prober, David
Keywords: none
MeSH Terms:
  • Animals
  • CpG Islands
  • Deoxyribonucleases/metabolism*
  • Germ-Line Mutation
  • INDEL Mutation
  • Mutagenesis*
  • Mutation
  • Zebrafish/embryology
  • Zebrafish/genetics
  • Zinc Fingers*
PubMed: 23303782 Full text @ Nucleic Acids Res.
ABSTRACT

Zinc-finger nucleases (ZFNs) and TAL effector nucleases (TALENs) have been shown to induce targeted mutations, but they have not been extensively tested in any animal model. Here, we describe a large-scale comparison of ZFN and TALEN mutagenicity in zebrafish. Using deep sequencing, we found that TALENs are significantly more likely to be mutagenic and induce an average of 10-fold more mutations than ZFNs. We observed a strong correlation between somatic and germ-line mutagenicity, and identified germ line mutations using ZFNs whose somatic mutations rates are well below the commonly used threshold of 1%. Guidelines that have previously been proposed to predict optimal ZFN and TALEN target sites did not predict mutagenicity in vivo. However, we observed a significant negative correlation between TALEN mutagenicity and the number of CpG repeats in TALEN target sites, suggesting that target site methylation may explain the poor mutagenicity of some TALENs in vivo. The higher mutation rates and ability to target essentially any sequence make TALENs the superior technology for targeted mutagenesis in zebrafish, and likely other animal models.

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