Targeted gene inactivation in zebrafish using engineered zinc-finger nucleases

Meng, Xiangdong; Noyes, Marcus B.; Zhu, Lihua Julie; Lawson, Nathan D.; Wolfe, Scot A.

doi:10.1038/nbt1398

Cited by 686 publications

(604 citation statements)

References 30 publications

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“…Here we show that Xenopus can be added to the growing list of important model animals for which ZFN-encoding mRNA has allowed facile reverse genetics, including Drosophila (7), zebrafish (22,27), and the rat (15,28). An important requirement for the use of ZFNs is a streamlined protocol to predict and implement effective gene disruption.…”

Section: Discussionmentioning

confidence: 99%

Efficient targeted gene disruption in the soma and germ line of the frog Xenopus tropicalis using engineered zinc-finger nucleases

Young

Cherone

Doyon

et al. 2011

Proc. Natl. Acad. Sci. U.S.A.

127

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The frog Xenopus , an important research organism in cell and developmental biology, currently lacks tools for targeted mutagenesis. Here, we address this problem by genome editing with zinc-finger nucleases (ZFNs). ZFNs directed against an eGFP transgene in Xenopus tropicalis induced mutations consistent with nonhomologous end joining at the target site, resulting in mosaic loss of the fluorescence phenotype at high frequencies. ZFNs directed against the noggin gene produced tadpoles and adult animals carrying up to 47% disrupted alleles, and founder animals yielded progeny carrying insertions and deletions in the noggin gene with no indication of off-target effects. Furthermore, functional tests demonstrated an allelic series of activity between three germ-line mutant alleles. Because ZFNs can be designed against any locus, our data provide a generally applicable protocol for gene disruption in Xenopus .

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Section: Discussionmentioning

confidence: 99%

Efficient targeted gene disruption in the soma and germ line of the frog Xenopus tropicalis using engineered zinc-finger nucleases

Young

Cherone

Doyon

et al. 2011

Proc. Natl. Acad. Sci. U.S.A.

127

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“…At the same time, genomic tools have enormously improved, facilitating the cloning of randomly induced mutations (Dahm and Geisler, 2006, Sivasubbu et al, 2007. In addition, methods of reverse genetics have been established, including gene knock-down via the injection of specific antisense morpholino oligos (MOs) (Nasevicius and Ekker, 2000, Doyon et al, 2008, Meng et al, 2008, Wienholds et al, 2003, target-selected ENU mutagenesis (also named TILLING; targeted induced local lesions in genomes) (Nasevicius and Ekker, 2000, Doyon et al, 2008, Meng et al, 2008, Wienholds et al, 2003, and, most recently, targeted introduction of small deletions via engineered zincfinger-nucleases (Nasevicius and Ekker, 2000, Doyon et al, 2008, Meng et al, 2008, Wienholds et al, 2003.…”

Section: Molecules and Genes With Essential Functions In Zebrafish Pimentioning

confidence: 99%

How to make a teleost adenohypophysis: Molecular pathways of pituitary development in zebrafish

Pogoda

Hammerschmidt

2009

Molecular and Cellular Endocrinology

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Please cite this article as: Pogoda, H.-M., Hammerschmidt, M., How to make a Teleost Adenohypophysis: Molecular Pathways of Pituitary development in Zebrafish, Molecular and Cellular Endocrinology (2008), doi:10.1016/j.mce.2009 This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. In the past years, the zebrafish has emerged as a powerful tool to elucidate the genetic networks controlling vertebrate development, behavior and disease. Based on mutants isolated in forward genetic screens and on gene knock-downs using morpholino oligonucleotide (oligo) antisense technology, our current understanding of the molecular machinery driving adenohypophyseal ontogeny could be considerably improved. In addition, comparative analyses have shed further light onto the evolution of this rather recently invented organ. The goal of this review is to summarize current knowledge of the genetic and molecular control of zebrafish pituitary development, with special focus on most recent findings, including some thus far unpublished data from our own laboratory on the transcription factor Six1. In addition, zebrafish data will be discussed in comparison with current understanding of adenohypophysis development in mouse.

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“…In many cases, ZFN-mediated DSBs have been used to stimulate the HR DNA-repair machinery for HR-mediated gene replacement and gene addition. This approach has been successfully implemented in animals (Beumer et al, 2006;Meng et al, 2008), human cell lines (Urnov et al, 2005;Lombardo et al, 2007), and plants (Wright et al, 2005;Cai et al, 2009;Shukla et al, 2009;Townsend et al, 2009;de Pater et al, 2013). However, it is important to note that while ZFNinduced DSBs can indeed induce the HR repair machinery, most of these breaks are repaired by the cell's NHEJ DNA-repair machinery in plant and other species.…”

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confidence: 99%

Nonhomologous End Joining-Mediated Gene Replacement in Plant Cells

2013

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Stimulation of the homologous recombination DNA-repair pathway via the induction of genomic double-strand breaks (DSBs) by zinc finger nucleases (ZFNs) has been deployed for gene replacement in plant cells. Nonhomologous end joining (NHEJ)-mediated repair of DSBs, on the other hand, has been utilized for the induction of site-specific mutagenesis in plants. Since NHEJ is the dominant DSB repair pathway and can also lead to the capture of foreign DNA molecules, we suggest that it can also be deployed for gene replacement. An acceptor DNA molecule in which a green fluorescent protein (GFP) coding sequence (gfp) was flanked by ZFN recognition sequences was used to produce transgenic target plants. A donor DNA molecule in which a promoterless hygromycin B phosphotransferase-encoding gene (hpt) was flanked by ZFN recognition sequences was constructed. The donor DNA molecule and ZFN expression cassette were delivered into target plants. ZFN-mediated sitespecific mutagenesis and complete removal of the GFP coding sequence resulted in the recovery of hygromycin-resistant plants that no longer expressed GFP and in which the hpt gene was unlinked to the acceptor DNA. More importantly, ZFNmediated digestion of both donor and acceptor DNA molecules resulted in NHEJ-mediated replacement of the gfp with hpt and recovery of hygromycin-resistant plants that no longer expressed GFP and in which the hpt gene was physically linked to the acceptor DNA. Sequence and phenotypical analyses, and transmission of the replacement events to the next generation, confirmed the stability of the NHEJ-induced gene exchange, suggesting its use as a novel method for transgene replacement and gene stacking in plants.

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Targeted gene inactivation in zebrafish using engineered zinc-finger nucleases

Cited by 686 publications

References 30 publications

Efficient targeted gene disruption in the soma and germ line of the frog Xenopus tropicalis using engineered zinc-finger nucleases

Efficient targeted gene disruption in the soma and germ line of the frog Xenopus tropicalis using engineered zinc-finger nucleases

How to make a teleost adenohypophysis: Molecular pathways of pituitary development in zebrafish

Nonhomologous End Joining-Mediated Gene Replacement in Plant Cells

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