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dc.contributor.authorAbu-Bonsrah, KD
dc.contributor.authorZhang, D
dc.contributor.authorNewgreen, DF
dc.date.accessioned2021-02-05T01:31:27Z
dc.date.available2021-02-05T01:31:27Z
dc.date.issued2016-10-03
dc.identifierpii: srep34524
dc.identifier.citationAbu-Bonsrah, K. D., Zhang, D. & Newgreen, D. F. (2016). CRISPR/Cas9 Targets Chicken Embryonic Somatic Cells In Vitro and In Vivo and generates Phenotypic Abnormalities. SCIENTIFIC REPORTS, 6 (1), https://doi.org/10.1038/srep34524.
dc.identifier.issn2045-2322
dc.identifier.urihttp://hdl.handle.net/11343/260429
dc.description.abstractChickens are an invaluable model for studying human diseases, physiology and especially development, but have lagged in genetic applications. With the advent of Programmable Engineered Nucleases, genetic manipulation has become efficient, specific and rapid. Here, we show that the CRISPR/Cas9 system can precisely edit the chicken genome. We generated HIRA, TYRP1, DICER, MBD3, EZH2, and 6 other gene knockouts in two chicken cell lines using the CRISPR/Cas9 system, with no off-target effects detected. We also showed that very large deletions (>75 kb) could be achieved. We also achieved targeted modification by homology-directed repair (HDR), producing MEN2A and MEN2B mutations of the RET gene. We also targeted DGCR8 in neural cells of the chicken embryo by in vivo electroporation. After FACS isolation of transfected cells, we observed appropriate sequence changes in DGCR8. Wholemount and frozen section antibody labelling showed reduction of DGCR8 levels in transfected cells. In addition, there was reduced expression levels of DGCR8-associated genes DROSHA, YPEL1 and NGN2. We also observed morphological differences in neural tissue and cardiac-related tissues of transfected embryos. These findings demonstrate that precisely targeted genetic manipulation of the genome using the CRISPR/Cas9 system can be extended to the highly adaptable in vivo chicken embryo model.
dc.languageEnglish
dc.publisherNATURE PUBLISHING GROUP
dc.rights.urihttps://creativecommons.org/licenses/by/4.0
dc.titleCRISPR/Cas9 Targets Chicken Embryonic Somatic Cells In Vitro and In Vivo and generates Phenotypic Abnormalities
dc.typeJournal Article
dc.identifier.doi10.1038/srep34524
melbourne.affiliation.departmentPaediatrics (RCH)
melbourne.affiliation.department
melbourne.affiliation.facultyMedicine, Dentistry & Health Sciences
melbourne.source.titleScientific Reports
melbourne.source.volume6
melbourne.source.issue1
dc.rights.licenseCC BY
melbourne.elementsid1104001
melbourne.contributor.authorNewgreen, Donald
melbourne.contributor.authorAbu-Bonsrah, Kwaku
melbourne.contributor.authorZhang, Dongcheng
dc.identifier.eissn2045-2322
melbourne.accessrightsOpen Access


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