Targeted mutagenesis of zebrafish: Use of zinc finger nucleases

Leong, Ivone; Lai, Daniel; Lan, Chuan-Ching; Johnson, Ross A.; Love, Donald R.

doi:10.1002/bdrc.20213

Cited by 11 publications

(9 citation statements)

References 65 publications

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“…Mutants are being generated using an ENU-mutagenesis-based method, TILLING [34], zinc finger nucleases (ZFNs) [5], and more recently, TALENs [4], which are proving to be efficacious in zebrafish [35]. Gene knockdowns using morpholinos is established [3], and with slight increases in efficiency, it may become possible to mutate a sufficient proportion of somatic cells to efficiently generate targeted knockouts by microinjections of fertilized eggs [35].…”

Section: Discussionmentioning

confidence: 99%

“…To begin to address this need, we have chosen the zebrafish as a model system because of the high degree of conservation of genes between fish and humans, the ability to score mutant phenotypes in the context of the organismal and tissue structure of the whole animal (especially embryonic phenotypes), and the growing ease of inhibiting the function of specific genes in zebrafish by antisense methods [3] or by mutation [4], [5]. The approach presented and referred to here as “ Hu manized Z ebrafish O rthologous R escue” (HuZOR) is based on the ability to rescue embryonic zebrafish mutant phenotypes by microinjection of mRNA into fertilized eggs.…”

Section: Introductionmentioning

confidence: 99%

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Functional Assessment of Human Coding Mutations Affecting Skin Pigmentation Using Zebrafish

et al. 2012

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A major challenge in personalized medicine is the lack of a standard way to define the functional significance of the numerous nonsynonymous, single nucleotide coding variants that are present in each human individual. To begin to address this problem, we have used pigmentation as a model polygenic trait, three common human polymorphisms thought to influence pigmentation, and the zebrafish as a model system. The approach is based on the rescue of embryonic zebrafish mutant phenotypes by “humanized” zebrafish orthologous mRNA. Two hypomorphic polymorphisms, L374F in SLC45A2, and A111T in SLC24A5, have been linked to lighter skin color in Europeans. The phenotypic effect of a second coding polymorphism in SLC45A2, E272K, is unclear. None of these polymorphisms had been tested in the context of a model organism. We have confirmed that zebrafish albino fish are mutant in slc45a2; wild-type slc45a2 mRNA rescued the albino mutant phenotype. Introduction of the L374F polymorphism into albino or the A111T polymorphism into slc24a5 (golden) abolished mRNA rescue of the respective mutant phenotypes, consistent with their known contributions to European skin color. In contrast, the E272K polymorphism had no effect on phenotypic rescue. The experimental conclusion that E272K is unlikely to affect pigmentation is consistent with a lack of correlation between this polymorphism and quantitatively measured skin color in 59 East Asian humans. A survey of mutations causing human oculocutaneous albinism yielded 257 missense mutations, 82% of which are theoretically testable in zebrafish. The developed approach may be extended to other model systems and may potentially contribute to our understanding the functional relationships between DNA sequence variation, human biology, and disease.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Functional Assessment of Human Coding Mutations Affecting Skin Pigmentation Using Zebrafish

et al. 2012

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“…Fish homozygous for rpl11 hi3820bTg , which was identified in a retroviral insertion mutagenesis screen, kept in standard conditions have been identified as models of Diamond-Blackfan anemia (Table 1A; Amsterdam et al 2004; Danilova et al 2014; Ear et al 2015; Y Zhang et al 2014). The improvement of reverse genetic techniques such as the use of zinc finger nucleases (Leong et al 2011; Sander et al 2011a, 2011b) CRISPRs (Hruscha and Schmid 2015; Hwang et al 2013; Hwang et al 2015) and transcription activator-like effector nucleases (TALENs; Huang et al 2011, 2016; Ma et al 2016; Sander et al 2011a, 2011b) have allowed for the targeted mutation of disease-specific genes. These techniques are becoming the method of choice in zebrafish for creating genetic models of human disease, such as models for visceral heterotaxy, Bethlem myopathy, and Allan-Herndon-Dudley syndrome (Table 1A; Noël et al 2015; Radev et al 2015; Zada et al 2014).…”

Section: Zfin As a Resource For Zebrafish Translational Researchmentioning

confidence: 99%

Zebrafish Models of Human Disease: Gaining Insight into Human Disease at ZFIN

et al. 2017

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The Zebrafish Model Organism Database (ZFIN; https://zfin.org) is the central resource for genetic, genomic, and phenotypic data for zebrafish (Danio rerio) research. ZFIN continuously assesses trends in zebrafish research, adding new data types and providing data repositories and tools that members of the research community can use to navigate data. The many research advantages and flexibility of manipulation of zebrafish have made them an increasingly attractive animal to model and study human disease.To facilitate disease-related research, ZFIN developed support to provide human disease information as well as annotation of zebrafish models of human disease. Human disease term pages at ZFIN provide information about disease names, synonyms, and references to other databases as well as a list of publications reporting studies of human diseases in which zebrafish were used. Zebrafish orthologs of human genes that are implicated in human disease etiology are routinely studied to provide an understanding of the molecular basis of disease. Therefore, a list of human genes involved in the disease with their corresponding zebrafish ortholog is displayed on the disease page, with links to additional information regarding the genes and existing mutations. Studying human disease often requires the use of models that recapitulate some or all of the pathologies observed in human diseases. Access to information regarding existing and published models can be critical, because they provide a tractable way to gain insight into the phenotypic outcomes of the disease. ZFIN annotates zebrafish models of human disease and supports retrieval of these published models by listing zebrafish models on the disease term page as well as by providing search interfaces and data download files to access the data. The improvements ZFIN has made to annotate, display, and search data related to human disease, especially zebrafish models for disease and disease-associated gene information, should be helpful to researchers and clinicians considering the use of zebrafish to study human disease.

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“…Zinc finger proteins, like transcription factors contain motifs that bind to DNA in a sequence specific manner. Therefore, upon expression, the zinc finger motif will bind to an endogenous target and the nuclease will induce double strand breaks in the DNA (Leong et al, 2011). The double strand brand breaks are generally repaired using non-homologous end joining, which is inherently error prone, thus leading to mutations.…”

Section: Zebrafish: a Missing Link?mentioning

confidence: 99%

The Necessity for In Vivo Functional Analysis in Human Medical Genetics.

Quintana

2015

MRAJ

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Approximately 50% of all congenital anomalies cannot be linked to any specific genetic etiology, but in recent years cost effective high throughput sequencing has emerged as an efficient strategy for identifying single nucleotide polymorphisms (SNPs) associated with disease. However, in many cases there is not enough evidence to determine if these SNPs underlie disease. To bridge this gap in our understanding advances in functional analyses are warranted. Several preclinical model systems are currently being utilized to provide such evidence, including the advantageous zebrafish embryo. While every system exhibits disadvantages and caveats, a new era of multidisciplinary research has evolved, which uses a broad spectrum of functional analysis tools. This approach will make it possible to identify potential therapeutic targets for both common and rare human disorders.

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Targeted mutagenesis of zebrafish: Use of zinc finger nucleases

Cited by 11 publications

References 65 publications

Functional Assessment of Human Coding Mutations Affecting Skin Pigmentation Using Zebrafish

Functional Assessment of Human Coding Mutations Affecting Skin Pigmentation Using Zebrafish

Zebrafish Models of Human Disease: Gaining Insight into Human Disease at ZFIN

The Necessity for In Vivo Functional Analysis in Human Medical Genetics.

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