title: [Construction of lentiviral expression vector containing Foxp3 gene and its expression in DC2.4 cell line].
abstract: OBJECTIVE
  To construct a Foxp3 lentiviral vector and transfer it into DC2.4 cells, which provides Foxp3+DC cells for further study on its immune modulation.
  
  
  METHODS
  We cloned mouse Foxp3 gene into lentiviral vector(pGC-FU) and acquired the plasmid pGC-FU-Foxp3. PCR and sequencing analysis were made for verifying the positive clones. The virus packaging plasmids(pGC-FU-Foxp3, pHelper1.0 and pHelper2.0) were contransfected into 293T cells, and the Lentivirus-Foxp3 was harvested from 293T cells. The Lentivirus-Foxp3 was used to infect DC2.4 cells in vitro and the expression of Foxp3 in infected DC2.4 cells was detected with flow cytometry(FCM).
  
  
  RESULTS
  PCR and sequencing revealed that the pGC-FU-Foxp3 plasmid was correctly constructed. The Lentivirus-Foxp3 with a titer of 2×10(8); TU/mL was successfully packaged. Foxp3 expression in DC2.4 cells infected with the Lentivirus-Foxp3 was increased significantly compared with negative control lentivirus.
  
  
  CONCLUSIONS
  The pGC-FU-Foxp3 plasmid has been successfully constructed and the Lentivirus-Foxp3 has been successfully packaged. Foxp3 can be enhanced in DC cells infected with the Lentivirus-Foxp3.
authors: Department of Ophthalmology, Chinese PLA General Hospital, Beijing 100853, China.
authors: Yu-bo
authors: YB
authors: Gong
authors: Yi-fei
authors: YF
authors: Huang
authors: Yan
authors: Y
authors: Li
authors: Gen-cheng
authors: GC
authors: Han
authors: Yong
authors: Y
authors: Liu
authors: Da-jiang
authors: DJ
authors: Wang
authors: Gai-ping
authors: GP
authors: DU
authors: Ji-feng
authors: JF
authors: Yu
mesh_terms: D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D003713:Dendritic Cells / Q000276:immunology / Q000378:metabolism*; D051858:Forkhead Transcription Factors / Q000235:genetics*; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics*; D056747:Immunomodulation; D016086:Lentivirus / Q000235:genetics*; D051379:Mice; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism
chemical_list: D051858:Forkhead Transcription Factors; C498833:Foxp3 protein, mouse; D012333:RNA, Messenger