title: High-efficiency and heritable gene targeting in mouse by transcription activator-like effector nucleases.
abstract: 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.
authors: Shanghai Key Laboratory of Regulatory Biology, Institute of Biomedical Sciences and School of Life Sciences, East China Normal University, Shanghai 200241, China.
authors: Zhongwei
authors: Z
authors: Qiu
authors: Meizhen
authors: M
authors: Liu
authors: Zhaohua
authors: Z
authors: Chen
authors: Yanjiao
authors: Y
authors: Shao
authors: Hongjie
authors: H
authors: Pan
authors: Gaigai
authors: G
authors: Wei
authors: Chao
authors: C
authors: Yu
authors: Long
authors: L
authors: Zhang
authors: Xia
authors: X
authors: Li
authors: Ping
authors: P
authors: Wang
authors: Heng-Yu
authors: HY
authors: Fan
authors: Bing
authors: B
authors: Du
authors: Bin
authors: B
authors: Liu
authors: Mingyao
authors: M
authors: Liu
authors: Dali
authors: D
authors: Li
mesh_terms: D000818:Animals; D004268:DNA-Binding Proteins / Q000235:genetics; D004706:Endodeoxyribonucleases / Q000235:genetics / Q000378:metabolism*; D018390:Gene Targeting / Q000379:methods*; D054643:INDEL Mutation; D051379:Mice; D059645:Mutation Rate; D010641:Phenotype
chemical_list: D004268:DNA-Binding Proteins; D004706:Endodeoxyribonucleases
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