Lentiviral vector-derived shRNAs confer enhanced suppression of Semliki forest virus replication in BHK-21 cells compared to shRNAs expressed from plasmids |
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Authors: | Ke Zhang Yu Chen Ji’an Pan Tero Ahola Deyin Guo |
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Institution: | (1) State Key Laboratory of Virology, The Modern Virology Research Centre, College of Life Sciences, Wuhan University, Wuhan, 430072, People’s Republic of China;(2) Institute of Biotechnology, Viikki Biocenter, University of Helsinki, 00014 Helsinki, Finland |
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Abstract: | Semliki forest virus (SFV) is a pathogen causing lethal encephalitis in laboratory mice. In this study, we obtained three
short hairpin RNAs (shRNAs) which could specifically target SFV sequence in GFP reporting systems and effectively suppress
SFV replication in luciferase-containing reporter virus system. At a multiplicity of infection (MOI) of 0.001, the luciferase
reporter activity was reduced by 78–92% by shRNA expression plasmids and virus yields reduced 2 to 10-fold at 20 h post-infection.
When lentiviral vector-derived shRNAs were employed, the virus titers decreased 8 to 126-fold at 24 h post-infection and 6
to 19-fold at 48 h post-infection and the cell survival was prolonged. These data formed the basis for further in vivo studies
of RNA interference in mouse models. |
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Keywords: | Lentiviral vector RNA interference Semliki forest virus Short hairpin RNA Small interfering RNA |
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