title: Synergistic inhibition of avian leukosis virus subgroup J replication by miRNA-embedded siRNA interference of double-target.
abstract: BACKGROUND
  The diseases caused by avian leukosis virus subgroup J (ALV-J) has become a serious problem in the poultry. Due to largely ineffective vaccines, new control measures are needed to be developed. RNA interference (RNAi) has been developed a promising measure for antivirus in poultry.
  
  
  METHODS
  In this study, miRNA-embedded siRNA interference was designed and used to inhibit ALV-J replication in vitro and in vivo. Each sequence of target siRNA derived from the gag (p15), pol (p32), env (gp85) and LTR (U3) gene of ALV-J was embedded into mouse miR-155 backbone as a pre-miRNA hairpin oligonucleotide sequence. After annealing, they were cloned into pcDNA6.2-GW/EmGFP-miR vector, respectively. For detecting the interference effect, recombinant vectors were introduced into DF-1 cells and day-old SPF chickens that infected with ALV-J.
  
  
  RESULTS
  In vitro, single target interference showed effective inhibition of reducing 74% ~ 85% mRNA of ALV-J. Double targets showed more efficient inhibition of reducing 96% ~ 98% mRNA of ALV-J. In vivo, chicks were inoculated with each recombinant plasmid in peritoneal cavity at day of hatch, and monitored infection status at interval 1 day postinfection for 4 weeks. Delivery of single target or double targets miRNA significantly reduced viremia and pathogenicity caused by ALV-J in vivo, especially the double targets.
  
  
  CONCLUSIONS
  These data demonstrated that the miRNA-embedded siRNA interference is an efficient method for inhibition of ALV-J replication, especially double targets.
authors.affiliation: College of Veterinary Medicine, Shandong Agricultural University, Tai'an, 271018, China. leng.lengaaa@163.com.
authors.forename: Rongrong
authors.initials: R
authors.lastname: Wei
authors.affiliation: College of Veterinary Medicine, Shandong Agricultural University, Tai'an, 271018, China. xiaoqianchen1014@163.com.
authors.forename: Xiaoqian
authors.initials: X
authors.lastname: Ma
authors.affiliation: College of Veterinary Medicine, Shandong Agricultural University, Tai'an, 271018, China. liujz@sdau.edu.cn.
authors.forename: Guihua
authors.initials: G
authors.lastname: Wang
authors.affiliation: College of Veterinary Medicine, Shandong Agricultural University, Tai'an, 271018, China. hjguo@sdau.edu.cn.
authors.forename: Huijun
authors.initials: H
authors.lastname: Guo
authors.affiliation: College of Veterinary Medicine, Shandong Agricultural University, Tai'an, 271018, China. liujz@sdau.edu.cn.
authors.forename: Jianzhu
authors.initials: J
authors.lastname: Liu
authors.affiliation: Xiangya School of Medicine, Changsha, 410013, China. gabrielle0731@gmail.com.
authors.forename: Lingxiao
authors.initials: L
authors.lastname: Fan
authors.affiliation: College of Veterinary Medicine, Shandong Agricultural University, Tai'an, 271018, China. czqsd@126.com.
authors.forename: Ziqiang
authors.initials: Z
authors.lastname: Cheng
mesh_terms: D000818:Animals; D001353:Avian Leukosis / Q000517:prevention & control / Q000821:virology*; D001354:Avian Leukosis Virus / Q000201:enzymology / Q000235:genetics* / Q000502:physiology; D002645:Chickens; D015536:Down-Regulation; D015683:Gene Products, gag / Q000235:genetics / Q000378:metabolism; D015684:Gene Products, pol / Q000235:genetics / Q000378:metabolism; D011201:Poultry Diseases / Q000517:prevention & control / Q000821:virology*; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics* / Q000378:metabolism; D020079:Terminal Repeat Sequences; D014779:Virus Replication
chemical_list: D015683:Gene Products, gag; D015684:Gene Products, pol; D034741:RNA, Small Interfering