Journal
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
Volume 109, Issue 43, Pages 17484-17489Publisher
NATL ACAD SCIENCES
DOI: 10.1073/pnas.1215421109
Keywords
genome editing; heritable mutagenesis; mutagenesis detection; reverse genetics; genome engineering
Categories
Funding
- Research Grants Council of Hong Kong [CUHK480709]
- National Basic Research Program of China [2009CB941202]
- Chinese Academy of Sciences [KSCX2-YW-R-083]
- National Institute of Child Health and Human Development, National Institutes of Health
- National Natural Science Foundation of China [31172394]
- Chinese University of Hong Kong
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Transcription activator-like effector nucleases (TALENs) are an approach for directed gene disruption and have been proved to be effective in various animal models. Here, we report that TALENs can induce somatic mutations in Xenopus embryos with reliably high efficiency and that such mutations are heritable through germ-line transmission. We modified the Golden Gate method for TALEN assembly to make the product suitable for RNA transcription and microinjection into Xenopus embryos. Eight pairs of TALENs were constructed to target eight Xenopus genes, and all resulted in indel mutations with high efficiencies of up to 95.7% at the targeted loci. Furthermore, mutations induced by TALENs were highly efficiently passed through the germ line to F-1 frogs. Together with simple and reliable PCR-based approaches for detecting TALEN-induced mutations, our results indicate that TALENs are an effective tool for targeted gene editing/knockout in Xenopus.
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