Please use this identifier to cite or link to this item: https://doi.org/10.1038/mt.2016.138
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dc.titleAdvanced design of dumbbell-shaped genetic minimal vectors improves non-coding and coding RNA expression
dc.contributor.authorJIANG XIAOOU
dc.contributor.authorYU HAN
dc.contributor.authorTEO CUI RONG
dc.contributor.authorTan, Genim Siu Xian
dc.contributor.authorGoh, Sok Chin
dc.contributor.authorPatel, Parasvi
dc.contributor.authorChua, Yiqiang Kevin
dc.contributor.authorHameed, Nasirah Banu Sahul
dc.contributor.authorAntonio Bertoletti
dc.contributor.authorVolker Patzel
dc.date.accessioned2018-01-31T02:48:11Z
dc.date.available2018-01-31T02:48:11Z
dc.date.issued2016-09-01
dc.identifier.citationJIANG XIAOOU, YU HAN, TEO CUI RONG, Tan, Genim Siu Xian, Goh, Sok Chin, Patel, Parasvi, Chua, Yiqiang Kevin, Hameed, Nasirah Banu Sahul, Antonio Bertoletti, Volker Patzel (2016-09-01). Advanced design of dumbbell-shaped genetic minimal vectors improves non-coding and coding RNA expression. Molecular Therapy 24 (9) : 1581-1591. ScholarBank@NUS Repository. https://doi.org/10.1038/mt.2016.138
dc.identifier.issn15250016
dc.identifier.issn15250024
dc.identifier.urihttp://scholarbank.nus.edu.sg/handle/10635/138636
dc.description.abstractDumbbell-shaped DNA minimal vectors lacking nontherapeutic genes and bacterial sequences are considered a stable, safe alternative to viral, nonviral, and naked plasmid-based gene-transfer systems. We investigated novel molecular features of dumbbell vectors aiming to reduce vector size and to improve the expression of noncoding or coding RNA. We minimized small hairpin RNA (shRNA) or microRNA (miRNA) expressing dumbbell vectors in size down to 130 bp generating the smallest genetic expression vectors reported. This was achieved by using a minimal H1 promoter with integrated transcriptional terminator transcribing the RNA hairpin structure around the dumbbell loop. Such vectors were generated with high conversion yields using a novel protocol. Minimized shRNA-expressing dumbbells showed accelerated kinetics of delivery and transcription leading to enhanced gene silencing in human tissue culture cells. In primary human T cells, minimized miRNA-expressing dumbbells revealed higher stability and triggered stronger target gene suppression as compared with plasmids and miRNA mimics. Dumbbell-driven gene expression was enhanced up to 56- or 160-fold by implementation of an intron and the SV40 enhancer compared with control dumbbells or plasmids. Advanced dumbbell vectors may represent one option to close the gap between durable expression that is achievable with integrating viral vectors and short-term effects triggered by naked RNA.
dc.language.isoen
dc.publisherNature Publishing Group
dc.subjectCell Line
dc.subjectEnhancer Elements, Genetic
dc.subjectGene Knockdown Techniques
dc.subjectGene Targeting
dc.subjectGenetic Vectors
dc.subjectHumans
dc.subjectIntrons
dc.subjectNucleic Acid Conformation
dc.subjectPlasmids
dc.subjectRNA Precursors
dc.subjectRNA Splicing
dc.subjectRNA, Messenger
dc.subjectRNA, Small Interfering
dc.subjectRNA, Untranslated
dc.subjectT-Lymphocytes
dc.subjectGene Expression
dc.typeArticle
dc.contributor.departmentDEPT OF MICROBIOLOGY & IMMUNOLOGY
dc.contributor.departmentDUKE-NUS GRADUATE MEDICAL SCHOOL S'PORE
dc.description.doi10.1038/mt.2016.138
dc.description.sourcetitleMolecular Therapy
dc.description.volume24
dc.description.issue9
dc.description.page1581-1591
dc.identifier.isiut000384962300010
dc.published.statePublished
dc.grant.idR-182-000-163-646
dc.grant.idNUHSRO/2015/091/Bridging/02
dc.grant.idNMRC/NIG/1058/2011
dc.grant.idT1-2011Sep-04 and T1-2014Apr-02
dc.grant.idT1-BSRG 2015-05
dc.grant.fundingagencyNational University of Singapore
dc.grant.fundingagencyNational Medical Research Council (Singapore)
dc.grant.fundingagencyMinistry of Education of Singapore
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