Please use this identifier to cite or link to this item: https://hdl.handle.net/10356/96391
Full metadata record
DC FieldValueLanguage
dc.contributor.authorYan, Yanen
dc.contributor.authorHong, Nien
dc.contributor.authorChen, Tianshengen
dc.contributor.authorLi, Mingyouen
dc.contributor.authorWang, Tiansuen
dc.contributor.authorGuan, Guijunen
dc.contributor.authorQiao, Yongkangen
dc.contributor.authorChen, Songlinen
dc.contributor.authorSchartl, Manfreden
dc.contributor.authorHong, Yunhanen
dc.contributor.authorLi, Chang Mingen
dc.date.accessioned2013-05-03T07:54:59Zen
dc.date.accessioned2019-12-06T19:29:50Z-
dc.date.available2013-05-03T07:54:59Zen
dc.date.available2019-12-06T19:29:50Z-
dc.date.copyright2013en
dc.date.issued2013en
dc.identifier.citationYan, Y., Hong, N., Chen, T., Li, M., Wang, T., Guan, G., Qiao, Y., Chen, S., Schartl, M., Li, C. M., & Hong, Y. (2013). p53 Gene Targeting by Homologous Recombination in Fish ES Cells. PLoS ONE, 8(3).en
dc.identifier.issn1932-6203en
dc.identifier.urihttps://hdl.handle.net/10356/96391-
dc.identifier.urihttp://hdl.handle.net/10220/9878en
dc.description.abstractGene targeting (GT) provides a powerful tool for the generation of precise genetic alterations in embryonic stem (ES) cells to elucidate gene function and create animal models for human diseases. This technology has, however, been limited to mouse and rat. We have previously established ES cell lines and procedures for gene transfer and selection for homologous recombination (HR) events in the fish medaka (Oryzias latipes). Methodology and Principal Findings: Here we report HR-mediated GT in this organism. We designed a GT vector to disrupt the tumor suppressor gene p53 (also known as tp53). We show that all the three medaka ES cell lines, MES1~MES3, are highly proficient for HR, as they produced detectable HR without drug selection. Furthermore, the positive-negative selection (PNS) procedure enhanced HR by ~12 folds. Out of 39 PNS-resistant colonies analyzed, 19 (48.7%) were positive for GT by PCR genotyping. When 11 of the PCR-positive colonies were further analyzed, 6 (54.5%) were found to be bona fide homologous recombinants by Southern blot analysis, sequencing and fluorescent in situ hybridization. This produces a high efficiency of up to 26.6% for p53 GT under PNS conditions. We show that p53 disruption and long-term propagation under drug selection conditions do not compromise the pluripotency, as p53-targeted ES cells retained stable growth, undifferentiated phenotype, pluripotency gene expression profile and differentiation potential in vitro and in vivo. Conclusions: Our results demonstrate that medaka ES cells are proficient for HR-mediated GT, offering a first model organism of lower vertebrates towards the development of full ES cell-based GT technology.en
dc.language.isoenen
dc.relation.ispartofseriesPLoS ONEen
dc.rights© 2013 The Author(s).en
dc.subjectDRNTU::Science::Biological sciences::Microbiology::Microorganismsen
dc.titleP53 gene targeting by homologous recombination in fish ES cellsen
dc.typeJournal Articleen
dc.contributor.schoolSchool of Chemical and Biomedical Engineeringen
dc.identifier.doi10.1371/journal.pone.0059400en
dc.description.versionPublished versionen
item.fulltextWith Fulltext-
item.grantfulltextopen-
Appears in Collections:SCBE Journal Articles
Files in This Item:
File Description SizeFormat 
4. p53 Gene Targeting by Homologous Recombination in Fish ES Cells.pdf18.5 MBAdobe PDFThumbnail
View/Open

Google ScholarTM

Check

Altmetric


Plumx

Items in DR-NTU are protected by copyright, with all rights reserved, unless otherwise indicated.