Abstract |
A robust artificial microRNA (amiRNA) strategy against porcine reproductive and respiratory syndrome virus (PRRSV) was developed by targeting the untranslated regions ( UTRs). Six candidate amiRNAs targeting the 5' or 3' UTR were used for vector construction, and four effective amiRNAs were selected for further study using a vector transfection/ virus infection assay. In cell cultures stably transfected with the four amiRNA vectors, expression of the sequence-specific amiRNAs was confirmed using poly(A)-tailed RT-PCR. After infection with three different PRRSV strains, the viral RNA genome and/or transcript were inhibited by ~90 % (semi-quantitative RT-PCR), and the viral titers were decreased by more than six log CCID(50) (viral titration assay) before day 3 postinfection. The potent anti-PRRSV effects lasted for at least 5 days. Sequence analysis showed that the amiRNA antiviral activities were not compromised by the presence of one or two mismatches in their binding targets. This work constitutes a step towards developing a more effective RNAi strategy against PRRSV.
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Authors | Bing Xia, Hongqin Song, Yang Chen, Xinyu Zhang, Xiaoli Xia, Huaichang Sun |
Journal | Archives of virology
(Arch Virol)
Vol. 158
Issue 1
Pg. 55-61
(Jan 2013)
ISSN: 1432-8798 [Electronic] Austria |
PMID | 22948796
(Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
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Chemical References |
- MicroRNAs
- RNA, Viral
- Untranslated Regions
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Topics |
- Animals
- Base Sequence
- Cell Line
- Down-Regulation
- MicroRNAs
(genetics, metabolism)
- Molecular Sequence Data
- Porcine Reproductive and Respiratory Syndrome
(virology)
- Porcine respiratory and reproductive syndrome virus
(genetics, physiology)
- RNA Interference
- RNA, Viral
(genetics, metabolism)
- Swine
- Untranslated Regions
- Virus Replication
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