Establishment and Evaluation of Stable Cell Lines Inhibiting Foot-and-Mouth Disease Virus by RNA Interference

RNA interference (RNAi) has been proved to be a powerful tool for foot-and-mouth disease virus FMDV inhibition in vitro and in vivo. We established five stable baby hamster kidney 21 cell lines (BHK-21) containing five short hairpin RNAs (shRNAs) expression plasmids (p3D1shRNA, p3D2shRNA, p3D3shRNA,...

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Bibliographic Details
Published in:BioMed research international 2014-01, Vol.2014 (2014), p.1-7
Main Authors: Zhang, Jie, Gao, Zong-liang, Zhou, Jian-hua, Liu, Yong-sheng, Gu, Yuan-xing
Format: Article
Language:English
Subjects:
Animals
Cell Culture Techniques - methods
Cell Line - virology
Cell lines
Cloning
Cricetinae
Deoxyribonucleic acid
DNA
Fluorescent Antibody Technique
Foot & mouth disease
Foot-and-mouth disease virus
Foot-and-Mouth Disease Virus - physiology
Genes
Genetic aspects
Genetic Vectors - metabolism
Genomes
Health aspects
Plasmids
Polymerase Chain Reaction
Real-Time Polymerase Chain Reaction
RNA Interference
RNA polymerase
RNA-protein interactions
Rodents
Science
Serial Passage
Transfection
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Summary:RNA interference (RNAi) has been proved to be a powerful tool for foot-and-mouth disease virus FMDV inhibition in vitro and in vivo. We established five stable baby hamster kidney 21 cell lines (BHK-21) containing five short hairpin RNAs (shRNAs) expression plasmids (p3D1shRNA, p3D2shRNA, p3D3shRNA, p3D4shRNA, and p3D5shRNA) targeting 3D gene of FMDV. Immunofluorescent assay, virus titration, and real-time quantitative reverse transcription polymerase chain reaction (Q-RT-PCR) were conducted to detect the effect of shRNAs on FMDV replication. After challenged with FMDV of O/CHA/99, two cell lines (p3D1shRNA and p3D4shRNA) showed a significant reduction in the synthesis of viral protein and RNA, accompanied by a sharp decrease in viral yield, and the inhibition could last for at least thirty passages. We developed an efficient procedure for the establishment and evaluation of stable cell lines for anti-FMDV research based on RNAi technology, which can be a candidate method for anti-FMDV research.
ISSN:2314-6133
2314-6141
DOI:10.1155/2014/109428