Construction of ciliary neurotrophic factor and green fluorescence protein-transduced recombinant adenovirus

VernacularTitle:构建睫状神经营养因子和绿色荧光蛋白基因转导的重组腺病毒载体
Author: Zheng-Qi LU ¹ ; Xue-Qiang HU ; Cau-Sheng ZHU ; Ran-Yi LIU ; Bi-Jun HUANG ; Wen-Lin HUANG
Author Information

1. 中山大学附属第三医院
Keywords: Ciliary neurotrophic factor,human; Gene recombination; Adenovirus vector; Gene therapy
From: Chinese Journal of Neuromedicine 2008;7(12):1196-1199
CountryChina
Language:Chinese
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.