PMID- 21667252
OWN - NLM
STAT- MEDLINE
DCOM- 20120525
LR  - 20211020
IS  - 1573-4978 (Electronic)
IS  - 0301-4851 (Linking)
VI  - 39
IP  - 3
DP  - 2012 Mar
TI  - Inhibition of porcine reproductive and respiratory syndrome virus replication by 
      RNA interference in MARC-145 cells.
PG  - 2515-22
LID - 10.1007/s11033-011-1003-z [doi]
AB  - With the ultimate aim of producing an RNA interference-mediated transgenic pig 
      that is resistant to porcine reproductive and respiratory syndrome virus (PRRSV), 
      we have investigated the effect of RNA interference (RNAi) on silencing the 
      expression of viral genes in the MARC-145 cell line. Twenty small interfering 
      RNAs (siRNAs) were designed and screened for their ability to suppress the 
      expression of the genes ORF1b, 5, 6, and 7 from the highly virulent isolate, 
      PRRSV-JXwn06. Of these siRNAs, the four most effective were selected and four 
      short hairpin RNA (shRNA) expression vectors (pGenesil-1-1b-135, 
      pGenesil-1-1b-372, pGenesil-1-6-135, and pGenesil-1-6-169) targeting ORF1b and 
      ORF6 were constructed and delivered into MARC-145 cells. These cells were then 
      infected with JXwn06. All four vectors inhibited the PRRSV-specific cytopathic 
      effect (CPE). The virus titers in cells transfected with pGenesil-1-1b-135, 
      pGenesil-1-1b-372, pGenesil-1-6-135, and pGenesil-1-6-169 were lower than that of 
      control cells by approximately 150-, 600-, 2.3- and 1.7-fold, respectively. In 
      addition, the expression levels of ORF1 and ORF6 were reduced compared with 
      controls. The unglycosylated membrane protein M, encoded by ORF6, was not 
      detectable in cells transfected with shRNA expression vectors. These results 
      verified that RNAi can effectively inhibit PRRSV-JXwn06 replication in cultured 
      cells in vitro. The four shRNA expression vectors are an initial step in the 
      production of transgenic pigs with PRRSV resistance.
FAU - Bao, Yonghua
AU  - Bao Y
AD  - Department of Basic Immunology, Xinxiang Medical University, Jinsui Street, 
      Xinxiang 453003, People's Republic of China. baoyonghua2005@yahoo.com.cn
FAU - Guo, Yongchen
AU  - Guo Y
FAU - Zhang, Liying
AU  - Zhang L
FAU - Zhao, Zhihui
AU  - Zhao Z
FAU - Li, Ning
AU  - Li N
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20110612
PL  - Netherlands
TA  - Mol Biol Rep
JT  - Molecular biology reports
JID - 0403234
RN  - 0 (RNA, Small Interfering)
SB  - IM
MH  - Animals
MH  - Blotting, Western
MH  - Cell Line
MH  - Gene Expression Regulation, Viral/*genetics
MH  - Genes, Viral/*genetics
MH  - Genetic Vectors/genetics
MH  - Haplorhini
MH  - Porcine respiratory and reproductive syndrome virus/genetics/*physiology
MH  - *RNA Interference
MH  - RNA, Small Interfering/genetics
MH  - Real-Time Polymerase Chain Reaction
MH  - Reverse Transcriptase Polymerase Chain Reaction
MH  - Transfection
MH  - Virus Replication/*genetics
EDAT- 2011/06/15 06:00
MHDA- 2012/05/26 06:00
CRDT- 2011/06/14 06:00
PHST- 2011/01/30 00:00 [received]
PHST- 2011/06/01 00:00 [accepted]
PHST- 2011/06/14 06:00 [entrez]
PHST- 2011/06/15 06:00 [pubmed]
PHST- 2012/05/26 06:00 [medline]
AID - 10.1007/s11033-011-1003-z [doi]
PST - ppublish
SO  - Mol Biol Rep. 2012 Mar;39(3):2515-22. doi: 10.1007/s11033-011-1003-z. Epub 2011 
      Jun 12.