PMID- 23732571
OWN - NLM
STAT- MEDLINE
DCOM- 20131231
LR  - 20191210
IS  - 1872-9096 (Electronic)
IS  - 0166-3542 (Linking)
VI  - 99
IP  - 2
DP  - 2013 Aug
TI  - A possible strategy to produce pigs resistant to porcine reproductive and 
      respiratory syndrome virus.
PG  - 158-64
LID - S0166-3542(13)00141-1 [pii]
LID - 10.1016/j.antiviral.2013.05.010 [doi]
AB  - The purpose of this study was to enhance the production of transgenic cloned 
      embryos with porcine reproductive and respiratory syndrome virus (PRRSV) shRNA 
      expression cassettes. To construct transgenic vector with expression targeting 
      against PRRSV, PRRSV shRNA expression cassettes were inserted into pEGFP-N1 and 
      the ability of resulting recombinant plasmid pEGFP-G1 inhibiting virus 
      replication was examined in Marc-145 cells. Results showed that PRRSV replication 
      could be significantly inhibited by pEGFP-G1 in Marc-145 cells compared with the 
      control. The pEGFP-G1 plasmid was used to deliver a transgene expressing EGFP and 
      the PRRSV shRNA into porcine fetal fibroblasts (PFF). Fluorescent-positive cells 
      were used as nuclear donors for somatic cell nuclear transfer (SCNT) to produce 
      shRNA-EGFP transgenic cloned embryos. The effects of trichostatin A (TSA) on 
      production of transgenic cloned embryos were investigated. Reconstructed embryos 
      were designed into 4 groups: Donor cells of Group A were treated with 50nM TSA 
      for 24h before SCNT. Reconstructed embryos of Group B were treated with 50nM TSA 
      for 24h after activation. Both donor cells and reconstructed embryos in Group C 
      were treated with TSA and Group D were the control without TSA treatment. The 
      results showed no difference (p>0.05) in cleavage rates among the 4 groups; 
      however, blastocyst developmental rates of Group B and C (30.9% and 42.0%, 
      respectively) were higher than for Group A and D (21.2% and 22.1%, respectively) 
      with Group C highest among groups (p<0.05). Interestingly, EGFP expression 
      intensity of transgenic cloned blastocysts of Group A was the highest. Our 
      results provide promising evidence toward a new approach for production of 
      transgenic cloned pigs with resistance to PRRSV and possibly a wide variety of 
      other porcine diseases.
CI  - Copyright (c) 2013 Elsevier B.V. All rights reserved.
FAU - Luo, Biping
AU  - Luo B
AD  - College of Veterinary Medicine, Nanjing Agricultural University, Jiangsu 210095, 
      China.
FAU - Ju, Shiqiang
AU  - Ju S
FAU - Wang, Bin
AU  - Wang B
FAU - Rui, Rong
AU  - Rui R
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20130602
PL  - Netherlands
TA  - Antiviral Res
JT  - Antiviral research
JID - 8109699
RN  - 0 (Hydroxamic Acids)
RN  - 0 (Protein Synthesis Inhibitors)
RN  - 0 (RNA, Small Interfering)
RN  - 0 (enhanced green fluorescent protein)
RN  - 147336-22-9 (Green Fluorescent Proteins)
RN  - 3X2S926L3Z (trichostatin A)
SB  - IM
MH  - Animals
MH  - Animals, Genetically Modified
MH  - Cell Line
MH  - Chlorocebus aethiops
MH  - Cloning, Organism
MH  - Embryo, Mammalian
MH  - Green Fluorescent Proteins/genetics
MH  - Hydroxamic Acids/pharmacology
MH  - Nuclear Transfer Techniques
MH  - Porcine Reproductive and Respiratory Syndrome/*virology
MH  - Porcine respiratory and reproductive syndrome virus/*genetics/*physiology
MH  - Protein Synthesis Inhibitors/pharmacology
MH  - RNA Interference
MH  - RNA, Small Interfering/biosynthesis/*genetics
MH  - Swine
MH  - Vero Cells
MH  - *Virus Replication
OTO - NOTNLM
OT  - Embryos
OT  - PRRSV
OT  - Pig
OT  - SCNT
OT  - TSA
OT  - shRNA
EDAT- 2013/06/05 06:00
MHDA- 2014/01/01 06:00
CRDT- 2013/06/05 06:00
PHST- 2013/03/25 00:00 [received]
PHST- 2013/05/21 00:00 [revised]
PHST- 2013/05/23 00:00 [accepted]
PHST- 2013/06/05 06:00 [entrez]
PHST- 2013/06/05 06:00 [pubmed]
PHST- 2014/01/01 06:00 [medline]
AID - S0166-3542(13)00141-1 [pii]
AID - 10.1016/j.antiviral.2013.05.010 [doi]
PST - ppublish
SO  - Antiviral Res. 2013 Aug;99(2):158-64. doi: 10.1016/j.antiviral.2013.05.010. Epub 
      2013 Jun 2.