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[Construction and expression of the self-inactivating lentiviral vector containing Murine Foxp3 gene].
- Source :
-
Sichuan da xue xue bao. Yi xue ban = Journal of Sichuan University. Medical science edition [Sichuan Da Xue Xue Bao Yi Xue Ban] 2009 Mar; Vol. 40 (2), pp. 199-202. - Publication Year :
- 2009
-
Abstract
- Objective: To construct the self-inactivating lentiviral vector containing murine Foxp3 gene and detect its expression in the CD4+ CD25- T cells.<br />Methods: The bicistronic lentiviral transfer plasmid containing Foxp3 gene and internal ribosomal entry site-green fluorescent protein gene (IRES-GFP) was constructed. Human embryonic kidney 293T cells were co-transfected by lentiviral packing system, which including the packaging plasmid deltaNRF, the transfer plasmid and the envelope plasmid VSVG using liposome. Then manifested the efficiency of gene transduction with Flow Cytometry (FCM) and the expressions of Foxp3 mRNA and protein were assayed by RT-PCR and Western blotting.<br />Results: The lentiviral transfer plasmid pXZ208-Foxp3-IRES-GFP was constructed, the virus titres were above 10(6) IU/mL in the supernatant. After coculture with thus cells, the CD4+ CD25- T cells in experimental group expressed significantly higher amounts of Foxp3 mRNA and protein than control group, and the efficiency of gene transduction was about 55.95%.<br />Conclusions: The self-inactivating lentiviral vector containing murine Foxp3 gene are successfully constructed. This gene delivery system can transduce Foxp3 gene into CD4+ CD25- T cells with highly efficient expression.
- Subjects :
- Animals
Forkhead Transcription Factors genetics
Green Fluorescent Proteins genetics
Green Fluorescent Proteins metabolism
Lentivirus metabolism
Mice
Mice, Inbred BALB C
RNA, Messenger genetics
RNA, Messenger metabolism
Forkhead Transcription Factors metabolism
Genetic Vectors
Lentivirus genetics
T-Lymphocytes, Regulatory metabolism
Transduction, Genetic
Subjects
Details
- Language :
- Chinese
- ISSN :
- 1672-173X
- Volume :
- 40
- Issue :
- 2
- Database :
- MEDLINE
- Journal :
- Sichuan da xue xue bao. Yi xue ban = Journal of Sichuan University. Medical science edition
- Publication Type :
- Academic Journal
- Accession number :
- 19462889