In vitro expression of hemophilia B gene mediated by lentivirus.
- Author:
Dong-Mei YAN
1
;
Kai-Lin XU
;
Bing DU
;
Ling-Yu ZENG
;
Qun-Xian LU
;
Xiu-Ying PAN
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Bone Marrow Cells; metabolism; Cells, Cultured; Dogs; Factor IX; genetics; metabolism; Genetic Vectors; Hemophilia B; genetics; metabolism; Humans; Lentivirus; genetics; Plasmids; genetics; Transfection
- From: Chinese Journal of Hematology 2008;29(9):583-586
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo construct a three plasmids lentiviral vector containing canine coagulation factor IX (cFIX) gene with ubiquinone promoter (PUB) and observe the expression of cFIX gene.
METHODSLentivirus was generated by transient three-plasmid transfection, namely, the VSV-G envelope expression cassette, the delta NRF packaging plasmid and the PTK 164 plasmid. Viral particles were used to infect the target cell, third passage mesenchymal stem cells (MSCs) and 293T cell respectively at MOI 3: 1. The cFIX activity was detected in cultured cells with one-stage clotting assay.
RESULTSThe MSCs were obtained in vitro. The lentivirus infected MSCs and 293T cells all expressed the active factor IX with the activity of (26.30 +/- 2.10)% and (19.70 +/- 1.53)%, respectively, which are significantly higher than that of control (1.00 +/- 0.05)%.
CONCLUSIONSThe lentiviral vector of three plasmids with ubiquinone promoter (PUB) was constructed and can transfect the MSCs and 293T cells.
