Please use this identifier to cite or link to this item: https://doi.org/10.1038/mt.2016.138
Title: Advanced design of dumbbell-shaped genetic minimal vectors improves non-coding and coding RNA expression
Authors: JIANG 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 
Keywords: Cell Line
Enhancer Elements, Genetic
Gene Knockdown Techniques
Gene Targeting
Genetic Vectors
Humans
Introns
Nucleic Acid Conformation
Plasmids
RNA Precursors
RNA Splicing
RNA, Messenger
RNA, Small Interfering
RNA, Untranslated
T-Lymphocytes
Gene Expression
Issue Date: 1-Sep-2016
Publisher: Nature Publishing Group
Citation: JIANG 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
Abstract: Dumbbell-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.
Source Title: Molecular Therapy
URI: http://scholarbank.nus.edu.sg/handle/10635/138636
ISSN: 15250016
15250024
DOI: 10.1038/mt.2016.138
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