RNAi-Mediated Gene silencing in Zebrafish Triggered by Convergent Transcription

Andrews, Omozusi E.; Diana, Jean-Sébastien; Wei, Chunyao; Patton, James G.

doi:10.1038/srep05222

Cited by 18 publications

(11 citation statements)

References 44 publications

(61 reference statements)

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“…In mouse oocytes, Dicer-dependent degradation of RNA pairs is thought to be essential for the proper regulation of female meiosis ( 9 , 10 ). Hence, this conserved RNAi-dependent mechanism can be used to artificially target silencing of genes in fission yeast and mammalian cells ( 7 ) as well as in zebrafish ( 8 ). It is therefore likely that the biological significance of asRNA molecules has not yet been fully unraveled.…”

Section: Discussionmentioning

confidence: 99%

“…On the other hand, it has long been known that convergent transcription of pericentromeric repeats in Schizosaccaromices pombe leads to the formation of heterochromatin and silencing ( 4 , 5 ). Converging transcript can indeed be used as a tool for transcriptional silencing not only in fission yeast but also zebrafish as well as mammalian cells ( 6 – 8 ). Also, compelling evidence in mouse oocytes has proven that antisense (as)RNA molecules can regulate genes and transposons at the post-transcriptional level both in cis and in trans ( 9 , 10 ).…”

Section: Introductionmentioning

confidence: 99%

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Post-transcriptional gene silencing triggered by sense transgenes involves uncapped antisense RNA and differs from silencing intentionally triggered by antisense transgenes

et al. 2015

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Although post-transcriptional gene silencing (PTGS) has been studied for more than a decade, there is still a gap in our understanding of how de novo silencing is initiated against genetic elements that are not supposed to produce double-stranded (ds)RNA. Given the pervasive transcription occurring throughout eukaryote genomes, we tested the hypothesis that unintended transcription could produce antisense (as)RNA molecules that participate to the initiation of PTGS triggered by sense transgenes (S-PTGS). Our results reveal a higher level of asRNA in Arabidopsis thaliana lines that spontaneously trigger S-PTGS than in lines that do not. However, PTGS triggered by antisense transgenes (AS-PTGS) differs from S-PTGS. In particular, a hypomorphic ago1 mutation that suppresses S-PTGS prevents the degradation of asRNA but not sense RNA during AS-PTGS, suggesting a different treatment of coding and non-coding RNA by AGO1, likely because of AGO1 association to polysomes. Moreover, the intended asRNA produced during AS-PTGS is capped whereas the asRNA produced during S-PTGS derives from 3′ maturation of a read-through transcript and is uncapped. Thus, we propose that uncapped asRNA corresponds to the aberrant RNA molecule that is converted to dsRNA by RNA-DEPENDENT RNA POLYMERASE 6 in siRNA-bodies to initiate S-PTGS, whereas capped asRNA must anneal with sense RNA to produce dsRNA that initiate AS-PTGS.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Post-transcriptional gene silencing triggered by sense transgenes involves uncapped antisense RNA and differs from silencing intentionally triggered by antisense transgenes

et al. 2015

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“…12 Recently, RNA-interference mediated chromatin silencing has been used to induce sequence-specific gene knockdown by convergent transcription (in which a DNA sequence is simultaneously transcribed in sense and anti-sense orientations directed by two inducible promoters) in zebrafish. 13 The targeted introduction of mutations using sequencespecific TALENs or the CRISPR system have recently been successfully applied to generate loss-of-function alleles by specifically targeting open reading frames or deletion or inversion of whole-chromosomal regions in vivo with efficiencies in zebrafish similar to those obtained using zinc-finger nucleases and transcriptionlike nucleases. 14 TALENs comprise a non-specific DNA-cleaving nuclease fused to an engineered DNA-binding domain, which contains a series of tandem repeats.…”

mentioning

confidence: 99%

The zebrafish eye—a paradigm for investigating human ocular genetics

Richardson¹,

Tracey‐White²,

Webster

et al. 2016

Eye

147

162

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Although human epidemiological and genetic studies are essential to elucidate the aetiology of normal and aberrant ocular development, animal models have provided us with an understanding of the pathogenesis of multiple developmental ocular malformations. Zebrafish eye development displays in depth molecular complexity and stringent spatiotemporal regulation that incorporates developmental contributions of the surface ectoderm, neuroectoderm and head mesenchyme, similar to that seen in humans. For this reason, and due to its genetic tractability, external fertilisation, and early optical clarity, the zebrafish has become an invaluable vertebrate system to investigate human ocular development and disease. Recently, zebrafish have been at the leading edge of preclinical therapy development, with their amenability to genetic manipulation facilitating the generation of robust ocular disease models required for large-scale genetic and drug screening programmes. This review presents an overview of human and zebrafish ocular development, genetic methodologies employed for zebrafish mutagenesis, relevant models of ocular disease, and finally therapeutic approaches, which may have translational leads in the future.

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“…MOs are especially useful to knockdown multiple alleles and when large gene families are genetically redundant [ 39 ]. Interference RNA is another popular technique for gene knockdown, but in strong contrast to Drosophila , C. elegans , and mammalian cells, few successful applications of shRNA [ 40 , 41 ] and iRNA [ 42 ] have been reported in zebrafish to silence gene expression. Despite concerted efforts in the field, progress in the use of iRNA technologies in zebrafish has been extremely slow.…”

Section: Mutagenesis and Knockdown In Zebrafish Before Crisprmentioning

confidence: 99%

Modeling Neuronal Diseases in Zebrafish in the Era of CRISPR

Espino-Saldaña¹,

Rodríguez-Ortiz

Pereida‐Jaramillo³

et al. 2020

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Background: Danio rerio is a powerful experimental model for studies in genetics and development. Recently, CRISPR technology has been applied in this species to mimic various human diseases, including those affecting the nervous system. Zebrafish offer multiple experimental advantages: external embryogenesis, rapid development, transparent embryos, short life cycle, and basic neurobiological processes shared with humans. This animal model, together with the CRISPR system, emerging imaging technologies, and novel behavioral approaches, lay the basis for a prominent future in neuropathology and will undoubtedly accelerate our understanding of brain function and its disorders. Objective: Gather relevant findings from studies that have used CRISPR technologies in zebrafish to explore basic neuronal function and model human diseases. Method: We systematically reviewed the most recent literature about CRISPR technology applications for understanding brain function and neurological disorders in D. rerio. We highlighted the key role of CRISPR in driving forward our understanding of particular topics in neuroscience. Results: We show specific advances in neurobiology when the CRISPR system has been applied in zebrafish and describe how CRISPR is accelerating our understanding of brain organization. Conclusion: Today, CRISPR is the preferred method to modify genomes of practically any living organism. Despite the rapid development of CRISPR technologies to generate disease models in zebrafish, more efforts are needed to efficiently combine different disciplines to find the etiology and treatments for many brain diseases.

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RNAi-Mediated Gene silencing in Zebrafish Triggered by Convergent Transcription

Cited by 18 publications

References 44 publications

Post-transcriptional gene silencing triggered by sense transgenes involves uncapped antisense RNA and differs from silencing intentionally triggered by antisense transgenes

Post-transcriptional gene silencing triggered by sense transgenes involves uncapped antisense RNA and differs from silencing intentionally triggered by antisense transgenes

The zebrafish eye—a paradigm for investigating human ocular genetics

Modeling Neuronal Diseases in Zebrafish in the Era of CRISPR

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