Please use this identifier to cite or link to this item: http://hdl.handle.net/1893/31524
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dc.contributor.authorJin, Ye Hwaen_UK
dc.contributor.authorLiao, Baoshanen_UK
dc.contributor.authorMigaud, Herveen_UK
dc.contributor.authorDavie, Andrewen_UK
dc.date.accessioned2020-08-06T00:02:49Z-
dc.date.available2020-08-06T00:02:49Z-
dc.date.issued2020-12en_UK
dc.identifier.other12600en_UK
dc.identifier.urihttp://hdl.handle.net/1893/31524-
dc.description.abstractThe application of genome engineering techniques to understand the mechanisms that regulate germ cell development opens promising new avenues to develop methods to control sexual maturation and mitigate associated detrimental effects in fish. In this study, the functional role of piwil2 in primordial germ cells (PGCs) was investigated in Nile tilapia using CRISPR/Cas9 and the resultant genotypes were further explored. piwil2 is a gonad-specific and maternally deposited gene in Nile tilapia eggs which is known to play a role in repression of transposon elements and is therefore thought to be important for maintaining germline cell fate. A functional domain of piwil2, PIWI domain, was targeted by injecting Cas9 mRNA and sgRNAs into Nile tilapia embryos at 1 cell stage. Results showed 54% of injected mutant larvae had no or less putative PGCs compared to control fish, suggesting an essential role of piwil2 in survival of PGCs. The genotypic features of the different phenotypic groups were explored by next generation sequencing (NGS) and other mutant screening methods including T7 endonuclease 1 (T7E1), CRISPR/Cas-derived RNA-guided engineered nuclease (RGEN), high resolution melt curve analysis (HRMA) and fragment analysis. Linking phenotypes to genotypes in F0 was hindered by the complex mosacism and wide indel spectrum revealed by NGS and fragment analysis. This study strongly suggests the functional importance of piwil2 in PGCs survival. Further studies should focus on reducing mosaicism when using CRISPR/Cas9 system to facilitate direct functional analysis in F0.en_UK
dc.language.isoenen_UK
dc.publisherSpringer Science and Business Media LLCen_UK
dc.relationJin YH, Liao B, Migaud H & Davie A (2020) Physiological impact and comparison of mutant screening methods in piwil2 KO founder Nile tilapia produced by CRISPR/Cas9 system. Scientific Reports, 10 (1), Art. No.: 12600. https://doi.org/10.1038/s41598-020-69421-0en_UK
dc.rightsThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.en_UK
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/en_UK
dc.subjectCRISPR-Cas9 genome editingen_UK
dc.subjectIchthyologyen_UK
dc.subjectReproductive biologyen_UK
dc.titlePhysiological impact and comparison of mutant screening methods in piwil2 KO founder Nile tilapia produced by CRISPR/Cas9 systemen_UK
dc.typeJournal Articleen_UK
dc.identifier.doi10.1038/s41598-020-69421-0en_UK
dc.identifier.pmid32724054en_UK
dc.citation.jtitleScientific Reportsen_UK
dc.citation.issn2045-2322en_UK
dc.citation.volume10en_UK
dc.citation.issue1en_UK
dc.citation.publicationstatusPublisheden_UK
dc.citation.peerreviewedRefereeden_UK
dc.type.statusVoR - Version of Recorden_UK
dc.contributor.funderSun Moon Universityen_UK
dc.contributor.funderUniversity of Stirlingen_UK
dc.citation.date28/07/2020en_UK
dc.contributor.affiliationInstitute of Aquacultureen_UK
dc.contributor.affiliationInstitute of Aquacultureen_UK
dc.contributor.affiliationInstitute of Aquacultureen_UK
dc.identifier.isiWOS:000556398500023en_UK
dc.identifier.scopusid2-s2.0-85088662663en_UK
dc.identifier.wtid1650348en_UK
dc.contributor.orcid0000-0002-5404-7512en_UK
dc.contributor.orcid0000-0002-9524-618Xen_UK
dc.date.accepted2020-07-09en_UK
dc.date.filedepositdate2020-08-05en_UK
Appears in Collections:Aquaculture Journal Articles

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