Knockout mice created by TALEN-mediated gene targeting

Sung, Young Hoon; Baek, In-Jeoung; Kim, Duk Hyoung; Jeon, Jisun; Lee, Jae‐Hoon; Lee, Kyung‐Hee; Jeong, Daewon; Kim, Jin-Soo; Lee, Han Woong

doi:10.1038/nbt.2477

Cited by 325 publications

(250 citation statements)

References 6 publications

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“…TALEN activity appeared to be dose-dependent, since injections of the higher dose (10 ng ml À 1 ) of Satb1 TALEN mRNA resulted in 5 of 11 (45.5%) F0 mice displaying TALEN-induced genomic modifications, while the lower concentration (5 ng ml À 1 ) yielded 12 of 56 (21.4%) F0 mice carrying a mutation at the TALENtargeting site (Table 2). In contrast to recently published data on the generation of TALEN-induced knockout mice 6 , we did not observe a clear difference in the percentage of mutant mice produced per oocyte injected for both concentrations of TALEN mRNA (5/297, 1.7% versus 12/426, 2.8%). This result speaks against significant off-target effects as also supported by our offtarget prediction (Supplementary Table 1).…”

Section: Generation Of a Targeting Vector For The Murine Satb1 Locuscontrasting

confidence: 99%

“…In line with recent findings, the majority of TALEN-induced mutations were deletions of variable length (Table 3) yielding 17 mutants (25.4%) with 4 mutant mice carrying bi-allelic mutations. In one mouse two differently mutated alleles and the wild-type allele were detected, suggesting a TALEN-induced chimerism in this animal (Table 3) 6 . TALEN activity appeared to be dose-dependent, since injections of the higher dose (10 ng ml À 1 ) of Satb1 TALEN mRNA resulted in 5 of 11 (45.5%) F0 mice displaying TALEN-induced genomic modifications, while the lower concentration (5 ng ml À 1 ) yielded 12 of 56 (21.4%) F0 mice carrying a mutation at the TALENtargeting site (Table 2).…”

Section: Generation Of a Targeting Vector For The Murine Satb1 Locusmentioning

confidence: 88%

“…This technology has been successfully applied in several animal models including mouse, rat, zebrafish and Caenorhabditis elegans [4][5][6][7][8] . Gene targeting via non-homologous end-joining (NHEJ) has also been suggested to generate combined reporter and knockout alleles 9 .…”

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confidence: 99%

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Efficient genome engineering by targeted homologous recombination in mouse embryos using transcription activator-like effector nucleases

et al. 2014

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Generation of mouse models by introducing transgenes using homologous recombination is critical for understanding fundamental biology and pathology of human diseases. Here we investigate whether artificial transcription activator-like effector nucleases (TALENs)-powerful tools that induce DNA double-strand breaks at specific genomic locations-can be combined with a targeting vector to induce homologous recombination for the introduction of a transgene in embryonic stem cells and fertilized murine oocytes. We describe the generation of a conditional mouse model using TALENs, which introduce double-strand breaks at the genomic locus of the special AT-rich sequence-binding protein-1 in combination with a large 14.4 kb targeting template vector. We report successful germline transmission of this allele and demonstrate its recombination in primary cells in the presence of Cre-recombinase. These results suggest that TALEN-assisted induction of DNA double-strand breaks can facilitate homologous recombination of complex targeting constructs directly in oocytes.

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Section: Generation Of a Targeting Vector For The Murine Satb1 Locuscontrasting

confidence: 99%

Section: Generation Of a Targeting Vector For The Murine Satb1 Locusmentioning

confidence: 88%

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Efficient genome engineering by targeted homologous recombination in mouse embryos using transcription activator-like effector nucleases

et al. 2014

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“…Zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and the clustered regularly interspaced short palindromic repeat (CRISPR)/Cas system are mainly used as engineered nucleases. A number of genetically modified animals have already been generated using such restriction enzymes (Bedell et al 2012;Geurts et al 2009;Hauschild et al 2011;Mashimo et al 2010;Ochiai et al 2010;Sung et al 2013;Suzuki et al 2013;Wang et al 2013;Yang et al 2013). Among these, the CRISPR/Cas system was developed the most recently and is particularly promising (Cong et al 2013;Mali et al 2013).…”

Section: Transgenic Techniques and Genome Editing Technology For Marmmentioning

confidence: 99%

Neuroscience Research Using Non-human Primate Models and Genome Editing

Kishi

2017

Research and Perspectives in Neurosciences

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The common marmoset (Callithrix jacchus) is a small New World non-human primate indigenous to northeastern Brazil. This species has been attracting the attention of biomedical researchers and neuroscientists for its ease of handling and colony maintenance, unique behavioral characteristics, and several human-like traits, such as enriched social vocal communication and strong relationships between parents and offspring. Its high reproductive efficiency makes it particularly amenable for use in the development of transgenic and genome editing technologies in a non-human primate model. Our group has recently generated transgenic marmosets with germ line transmission, opening new avenues in primate research.In this chapter, we describe recent advances in neuroscience and disease research using common marmosets, and we outline potential uses of genome editing in non-human primates toward the development of knock-in/knock-out marmosets.

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“…The TALENs have rapidly gained prominence as a novel genome-editing tool, which were successfully applied to create site-specific gene modifications in model organisms such as yeast, plants, zebra fish, mouse, rat and human cells, including human pluripotent cells. [58][59][60][61][62] TALEN has also been used to generate single base-pair mutations, linking single-nucleotide polymorphisms to specific human disease. 63 Furthermore, TALENs have even been utilized to eliminate the mutant form of mitochondrial DNA from patient-derived cells.…”

Section: Talenmentioning

confidence: 99%

A practical guide to induced pluripotent stem cell research using patient samples

Santostefano

Hamazaki

Biel

et al. 2015

Laboratory Investigation

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Approximately 3 years ago, we assessed how patient induced pluripotent stem cell (iPSC) research could potentially impact human pathobiology studies in the future. Since then, the field has grown considerably with numerous technical developments, and the idea of modeling diseases 'in a dish' is becoming increasingly popular in biomedical research. Likely, it is even acceptable to include patient iPSCs as one of the standard research tools for disease mechanism studies, just like knockout mice. However, as the field matures, we acknowledge there remain many practical limitations and obstacles for their genuine application to understand diseases, and accept that it has not been as straightforward to model disorders as initially proposed. A major practical challenge has been efficient direction of iPSC differentiation into desired lineages and preparation of the large numbers of specific cell types required for study. Another even larger obstacle is the limited value of in vitro outcomes, which often do not closely represent disease conditions. To overcome the latter issue, many new approaches are underway, including three-dimensional organoid cultures from iPSCs, xenotransplantation of human cells to animal models and in vitro interaction of multiple cell types derived from isogenic iPSCs. Here we summarize the areas where patient iPSC studies have provided truly valuable information beyond existing skepticism, discuss the desired technologies to overcome current limitations and include practical guidance for how to utilize the resources. Undoubtedly, these human patient cells are an asset for experimental pathology studies. The future rests on how wisely we use them.

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Knockout mice created by TALEN-mediated gene targeting

Cited by 325 publications

References 6 publications

Efficient genome engineering by targeted homologous recombination in mouse embryos using transcription activator-like effector nucleases

Efficient genome engineering by targeted homologous recombination in mouse embryos using transcription activator-like effector nucleases

Neuroscience Research Using Non-human Primate Models and Genome Editing

A practical guide to induced pluripotent stem cell research using patient samples

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