Construction of ciliary neurotrophic factor and green fluorescence protein-transduced recombinant adenovirus
- Author:
- No author available
- Journal Title:
- CHINESE JOURNAL OF NEUROMEDICINE
- Issue:
- 12
- DOI:
- 10.3760/cma.j.issn.1671-8925.2008.12.003
- Key Word:
- 睫状神经营养因子人源性;基因重组;腺病毒载体;基因治疗;Ciliary neurotrophic factor,human;Gene recombination;Adenovirus vector;Gene therapy
Abstract： Objective To construct an adenoviral vector carrying the gene encoding ciliary neurotrophic factor (CNTF). Methods The gene fragment encoding CNTF was amplified from pMEG-CNTF plasmid by PCR and the Psp-CNTF-IRES-GFP and PDC316-CNTF-IRES-GFP plasmids were constructed. Using PDC316-CNTF-IRES-GFP and PBHG plasmids, the Ad-CNTF-IRES-GFP vector was constructed, and the constructed vector was amplified, purified and identified in 293-LP cells. Ectopic overexpression of CNTF was induced using the constructed vector in human bone marrow-derived mesenchymal stem cells (MSCs) to investigate the role of CNTF in promoting remyelination. Results The Ad-CNTF-IRES-GFP vector was successfully constructed with a pfu of 2.3x1011. CNTF concentration in the MSCs transfeeted with Ad-CNTF-IRES-GFP vector was 20-fold higher than that in either non-transfected or Ad-EGFP-transfected MSCs. Conclusion The constructed Ad-CNTF-IRES-GFP vector allows CNTF overexpression in human MSCs by 20 folds, which provides a strategy for gene therapy targeting CNTF.
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