Journal Article

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

Zhongwei Qiu, Meizhen Liu, Zhaohua Chen, Yanjiao Shao, Hongjie Pan, Gaigai Wei, Chao Yu, Long Zhang, Xia Li, Ping Wang, Heng-Yu Fan, Bing Du, Bin Liu, Mingyao Liu and Dali Li

in Nucleic Acids Research

Volume 41, issue 11, pages e120-e120
Published in print June 2013 | ISSN: 0305-1048
Published online April 2013 | e-ISSN: 1362-4962 | DOI: https://dx.doi.org/10.1093/nar/gkt258

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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.

Journal Article.  5330 words.  Illustrated.

Subjects: Chemistry ; Biochemistry ; Bioinformatics and Computational Biology ; Genetics and Genomics ; Molecular and Cell Biology