title: [Construction and identification of mouse eosinophils CCR3 gene RNA interference lentiviral vector].
abstract: OBJECTIVE
  Through construction of a lentiviral expression vector of chemokine receptor 3 (CCR3)RNA interference (RNAi) of mouse, to further study the function of CCR3 gene on eosinophils.
  
  
  METHODS
  Focused on the CCR3 gene sequences, RNAi target sequences were designed, then the target sequences of Oligo DNA were synthesized and annealed to double stranded DNA, which was subsequently connected to pLVX-shRNA2-m vector digested by MluI, SacI, EcoRI, HindIII, BamHI and Xho I, short hairpin RNA lentiviral vectors were constructed. Short hairpin RNA lentiviral vectors were constructed. 293T cells and eosinophils were transfected by shRNA lentiviral vector, and virus titer was determined. The expression of the CCR3 gene in eosinophils was identified by quantitative-PCR.
  
  
  RESULTS
  The lentiviral vector of shRNA-mCCR3-oligonucleotide chain was inserted correctly. Infection efficiency of 293T cells observed under fluorescence microscope was more than 90%, the virus titer was 4×10(8) TU/ml. CCR3 interference rate was 86.7%.
  
  
  CONCLUSIONS
  A lentiviral vector of CCR3-gene RNAi was constructed successfully by the genetic engineering technology, and it provides a condition for further research in vitro and vivo.
authors.affiliation: Department of Otorhinolaryngology Head and Neck Surgery, Second Affiliated Hospital, Nanchang University, Nanchang 330006, China.
authors.forename: Xin-hua
authors.initials: XH
authors.lastname: Zhu
authors.forename: Bing
authors.initials: B
authors.lastname: Liao
authors.forename: Xin-yue
authors.initials: XY
authors.lastname: Wang
authors.forename: Ke
authors.initials: K
authors.lastname: Liu
authors.forename: Yue-hui
authors.initials: YH
authors.lastname: Liu
mesh_terms: D000818:Animals; D001483:Base Sequence; D004247:DNA; D004804:Eosinophils / Q000378:metabolism*; D005822:Genetic Vectors; D016086:Lentivirus / Q000235:genetics*; D051379:Mice; D016133:Polymerase Chain Reaction; D034622:RNA Interference; D034741:RNA, Small Interfering; D054397:Receptors, CCR3 / Q000235:genetics* / Q000378:metabolism; D014162:Transfection
chemical_list: C515351:Ccr3 protein, mouse; D034741:RNA, Small Interfering; D054397:Receptors, CCR3; D004247:DNA