CRISPR/Cas9-Mediated Knockin and Knockout in Zebrafish

Albadri, Shahad; Santis, Flavia De; Donato, Vincenzo Di; Bene, Filippo Del

doi:10.1007/978-3-319-60192-2_4

Cited by 21 publications

(14 citation statements)

References 38 publications

(39 reference statements)

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“…Additionally, adult social behaviors in a zebrafish ASD model have reduced shoaling behavior (Liu et al, 2018). While the bulk of current zebrafish ASD models are caused by indels that result in premature stop codons, recently developed CRISPR/Cas9 technologies in zebrafish are making tissue-specific mutagenesis and knockins that replicate patient-specific mis-sense mutations more broadly feasible (Albadri et al, 2017;. These technological advances continue to make the zebrafish an increasingly valuable model for ASD research.…”

Section: Discussion: Drawing Conclusion and Looking Forwardmentioning

confidence: 99%

The Gut-Brain-Microbiome Axis and Its Link to Autism: Emerging Insights and the Potential of Zebrafish Models

James

Davidson

Yanes

et al. 2021

Front. Cell Dev. Biol.

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Research involving autism spectrum disorder (ASD) most frequently focuses on its key diagnostic criteria: restricted interests and repetitive behaviors, altered sensory perception, and communication impairments. These core criteria, however, are often accompanied by numerous comorbidities, many of which result in severe negative impacts on quality of life, including seizures, epilepsy, sleep disturbance, hypotonia, and GI distress. While ASD is a clinically heterogeneous disorder, gastrointestinal (GI) distress is among the most prevalent co-occurring symptom complex, manifesting in upward of 70% of all individuals with ASD. Consistent with this high prevalence, over a dozen family foundations that represent genetically distinct, molecularly defined forms of ASD have identified GI symptoms as an understudied area with significant negative impacts on quality of life for both individuals and their caregivers. Moreover, GI symptoms are also correlated with more pronounced irritability, social withdrawal, stereotypy, hyperactivity, and sleep disturbances, suggesting that they may exacerbate the defining behavioral symptoms of ASD. Despite these facts (and to the detriment of the community), GI distress remains largely unaddressed by ASD research and is frequently regarded as a symptomatic outcome rather than a potential contributory factor to the behavioral symptoms. Allowing for examination of both ASD’s impact on the central nervous system (CNS) as well as its impact on the GI tract and the associated microbiome, the zebrafish has recently emerged as a powerful tool to study ASD. This is in no small part due to the advantages zebrafish present as a model system: their precocious development, their small transparent larval form, and their parallels with humans in genetics and physiology. While ASD research centered on the CNS has leveraged these advantages, there has been a critical lack of GI-centric ASD research in zebrafish models, making a holistic view of the gut-brain-microbiome axis incomplete. Similarly, high-throughput ASD drug screens have recently been developed but primarily focus on CNS and behavioral impacts while potential GI impacts have not been investigated. In this review, we aim to explore the great promise of the zebrafish model for elucidating the roles of the gut-brain-microbiome axis in ASD.

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Section: Discussion: Drawing Conclusion and Looking Forwardmentioning

confidence: 99%

The Gut-Brain-Microbiome Axis and Its Link to Autism: Emerging Insights and the Potential of Zebrafish Models

James

Davidson

Yanes

et al. 2021

Front. Cell Dev. Biol.

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“…The Clustered Regulatory Interspaced Short Palindromic Repeats (CRISPR) and CRISPR-associated (Cas) system has brought a remarkable development in genome engineering efficiency in different organisms during the past few years (Doudna and Charpentier, 2014;Albadri et al, 2017;Ermert et al, 2019;Pu et al, 2019;Song et al, 2019). Briefly, the CRISPR-cas system employs a guide RNA (gRNA) and an endonuclease, mostly a single nuclease Cas9 (Makarova et al, 2011;Chylinski et al, 2014), as the two working elements for a site directed DNA cutting.…”

Section: Crispr-casmentioning

confidence: 99%

The Gibberellin Producer Fusarium fujikuroi: Methods and Technologies in the Current Toolkit

Cen

Lin

Wang

et al. 2020

Front. Bioeng. Biotechnol.

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In recent years, there has been a noticeable increase in research interests on the Fusarium species, which includes prevalent plant pathogens and human pathogens, common microbial food contaminants and industrial microbes. Taken the advantage of gibberellin synthesis, Fusarium fujikuroi succeed in being a prevalent plant pathogen. At the meanwhile, F. fujikuroi was utilized for industrial production of gibberellins, a group of extensively applied phytohormone. F. fujikuroi has been known for its outstanding performance in gibberellin production for almost 100 years. Research activities relate to this species has lasted for a very long period. The slow development in biological investigation of F. fujikuroi is largely due to the lack of efficient research technologies and molecular tools. During the past decade, technologies to analyze the molecular basis of host-pathogen interactions and metabolic regulations have been developed rapidly, especially on the aspects of genetic manipulation. At the meanwhile, the industrial fermentation technologies kept sustained development. In this article, we reviewed the currently available research tools/methods for F. fujikuroi research, focusing on the topics about genetic engineering and gibberellin production.

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“…To this end, a vector system called 2C-Cas9 (Cre-mediated recombination for Clonal analysis of Cas9 mutant cells) was developed, based on the UAS-driven expression of Cas9 and U6-driven expression of two different sgRNAs. UAS-driven expression of Cas9 offers the possibility of conditional targeted mutagenesis in virtually any cell-type through the use of the broad repertoire of available tissue-specific zebrafish transgenic Gal4 driver lines ( Di Donato et al, 2016 ; Albadri et al, 2017a ).…”

Section: Crispr/cas9 In Zebrafishmentioning

confidence: 99%

Combining Zebrafish and CRISPR/Cas9: Toward a More Efficient Drug Discovery Pipeline

2018

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The use of zebrafish larvae in basic and applied research has grown exponentially during the last 20 years. The reasons for this success lay in its specific experimental advantages: on the one hand, the small size, the large number of progeny and the fast life cycle greatly facilitate large-scale approaches while maintaining 3Rs amenability; on the other hand, high genetic and physiological homology with humans and ease of genetic manipulation make zebrafish larvae a highly robust model for understanding human disease. Together, these advantages allow using zebrafish larvae for performing high-throughput research, both in terms of chemical and genetic phenotypic screenings. Therefore, the zebrafish larva as an animal model is placed between more reductionist in vitro high-throughput screenings and informative but low-throughput preclinical assays using mammals. However, despite its biological advantages and growing translational validation, zebrafish remains scarcely used in current drug discovery pipelines. In a context in which the pharmaceutical industry is facing a productivity crisis in bringing new drugs to the market, the combined advantages of zebrafish and the CRISPR/Cas9 system, the most powerful technology for genomic editing to date, has the potential to become a valuable tool for streamlining the generation of models mimicking human disease, the validation of novel drug targets and the discovery of new therapeutics. This review will focus on the most recent advances on CRISPR/Cas9 implementation in zebrafish and all their potential uses in biomedical research and drug discovery.

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CRISPR/Cas9-Mediated Knockin and Knockout in Zebrafish

Cited by 21 publications

References 38 publications

The Gut-Brain-Microbiome Axis and Its Link to Autism: Emerging Insights and the Potential of Zebrafish Models

The Gut-Brain-Microbiome Axis and Its Link to Autism: Emerging Insights and the Potential of Zebrafish Models

The Gibberellin Producer Fusarium fujikuroi: Methods and Technologies in the Current Toolkit

Combining Zebrafish and CRISPR/Cas9: Toward a More Efficient Drug Discovery Pipeline

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