High-efficiency and heritable gene targeting in mouse by transcription activator-like effector nucleases

Transcription activator-like effector nucleases (TALENs) are a powerful new approach for targeted gene disruption in various animal models, but little is known about their activities in Mus musculus, the widely used mammalian model organism. Here, we report that direct injection of in vitro transcri...

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Bibliographic Details
Published in:Nucleic acids research 2013-06, Vol.41 (11), p.e120-e120
Main Authors: Qiu, Zhongwei, Liu, Meizhen, Chen, Zhaohua, Shao, Yanjiao, Pan, Hongjie, Wei, Gaigai, Yu, Chao, Zhang, Long, Li, Xia, Wang, Ping, Fan, Heng-Yu, Du, Bing, Liu, Bin, Liu, Mingyao, Li, Dali
Format: Article
Language:English
Subjects:
Animals
DNA-Binding Proteins - genetics
Endodeoxyribonucleases - genetics
Endodeoxyribonucleases - metabolism
Gene Targeting - methods
INDEL Mutation
Methods Online
Mice
Mutation Rate
Phenotype
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Summary:Transcription activator-like effector nucleases (TALENs) are a powerful new approach for targeted gene disruption in various animal models, but little is known about their activities in Mus musculus, the widely used mammalian model organism. Here, we report that direct injection of in vitro transcribed messenger RNA of TALEN pairs into mouse zygotes induced somatic mutations, which were stably passed to the next generation through germ-line transmission. With one TALEN pair constructed for each of 10 target genes, mutant F0 mice for each gene were obtained with the mutation rate ranged from 13 to 67% and an average of ∼40% of total healthy newborns with no significant differences between C57BL/6 and FVB/N genetic background. One TALEN pair with single mismatch to their intended target sequence in each side failed to yield any mutation. Furthermore, highly efficient germ-line transmission was obtained, as all the F0 founders tested transmitted the mutations to F1 mice. In addition, we also observed that one bi-allele mutant founder of Lepr gene, encoding Leptin receptor, had similar diabetic phenotype as db/db mouse. Together, our results suggest that TALENs are an effective genetic tool for rapid gene disruption with high efficiency and heritability in mouse with distinct genetic background.
ISSN:0305-1048
1362-4962
DOI:10.1093/nar/gkt258