Please use this identifier to cite or link to this item: https://doi.org/10.1093/nar/gkv583
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dc.titleEfficient production of superior dumbbell-shaped DNA minimal vectors for small hairpin RNA expression
dc.contributor.authorYU HAN
dc.contributor.authorJIANG XIAOOU
dc.contributor.authorTAN KAR TONG
dc.contributor.authorHang, Liting
dc.contributor.authorVolker Patzel
dc.date.accessioned2018-01-31T09:08:56Z
dc.date.available2018-01-31T09:08:56Z
dc.date.issued2015-01-01
dc.identifier.citationYU HAN, JIANG XIAOOU, TAN KAR TONG, Hang, Liting, Volker Patzel (2015-01-01). Efficient production of superior dumbbell-shaped DNA minimal vectors for small hairpin RNA expression. Nucleic Acids Research 43 (18). ScholarBank@NUS Repository. https://doi.org/10.1093/nar/gkv583
dc.identifier.issn03051048
dc.identifier.issn13624962
dc.identifier.urihttp://scholarbank.nus.edu.sg/handle/10635/138642
dc.description.abstractGenetic therapy holds great promise for the treatment of inherited or acquired genetic diseases; however, its breakthrough is hampered by the lack of suitable gene delivery systems. Dumbbell-shaped DNA minimal vectors represent an attractive, safe alternative to the commonly used viral vectors which are fraught with risk, but dumbbell generation appears to be costly and time-consuming. We developed a new PCR-based method for dumbbell production which comprises only two steps. First, PCR amplification of the therapeutic expression cassette using chemically modified primers to form a ready-to-ligate DNA structure; and second, a highly efficient intramolecular ligation reaction. Compared with conventional strategies, the new method produces dumbbell vectors more rapidly, with higher yields and purity, and at lower costs. In addition, such produced small hairpin RNA expressing dumbbells triggered superior target gene knockdown compared with conventionally produced dumbbells or plasmids. Our novel method is suitable for large-scale dumbbell production and can facilitate clinical applications of this vector system.
dc.language.isoen
dc.publisherOxford University Press
dc.rightsAttribution-NonCommercial 4.0 International
dc.rights.urihttp://creativecommons.org/licenses/by-nc/4.0/
dc.subjectCell Line
dc.subjectDNA
dc.subjectDNA Primers
dc.subjectFurans
dc.subjectGene Expression
dc.subjectGene Knockout Techniques
dc.subjectGenetic Vectors
dc.subjectHumans
dc.subjectPolymerase Chain Reaction
dc.subjectRNA, Small Interfering
dc.typeArticle
dc.contributor.departmentCANCER SCIENCE INSTITUTE OF SINGAPORE
dc.contributor.departmentMICROBIOLOGY & IMMUNOLOGY
dc.description.doi10.1093/nar/gkv583
dc.description.sourcetitleNucleic Acids Research
dc.description.volume43
dc.description.issue18
dc.identifier.isiut000366406500006
dc.published.statePublished
dc.grant.idNIG/1058/2011
dc.grant.idT1–2011Sep-04] and [T1–2014Apr-02
dc.grant.fundingagencyNational University of Singapore
dc.grant.fundingagencyNational Medical Research Council (Singapore)
dc.grant.fundingagencyMinistry of Education (MOE)
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