(2016) Using intron splicing trick for preferential gene expression in transduced cells: an approach for suicide gene therapy. Cancer Gene Therapy. pp. 7-12. ISSN 0929-1903
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Abstract
Suicide gene therapy is one of the most innovative approaches in which a potential toxic gene is delivered to the targeted cancer cell by different target delivery methods. We constructed a transfer vector to express green fluorescent protein (GFP) in transduced cells but not in packaging cells. We placed gfp under the control of the cytomegalovirus (CMV) promoter, which is positioned between the two long-terminal repeats in reverse direction. The intron-2 sequence of the human beta globin gene with two poly-A signals and several stop codons on the antisense strand was placed on the leading strand between the CMV promoter and gfp. For lentiviral production, the HEK293T and line were co-transfected with the PMD2G, psPAX2 and pLentiGFP-Ins2 plasmids. The HEK293T and line were transduced with this virus. PCR was performed for evaluation of intron splicing in transduced cells. The GFP expression was seen in 65 of the cells transduced. The PCR amplification of the genomic DNA of transduced cells confirmed the splicing of intron 2. The strategy is significant to accomplish our goal for preserving the packaging cells from the toxic gene expression during viral assembly and the resultant reduction in viral titration. Also it serves to address several other issues in the gene therapy.
Item Type: | Article |
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Keywords: | diphtheria-toxin lentiviral vector toxicity systems chain |
Page Range: | pp. 7-12 |
Journal or Publication Title: | Cancer Gene Therapy |
Journal Index: | ISI |
Volume: | 23 |
Number: | 1 |
Identification Number: | https://doi.org/10.1038/cgt.2015.57 |
ISSN: | 0929-1903 |
Depositing User: | مهندس مهدی شریفی |
URI: | http://eprints.mui.ac.ir/id/eprint/3188 |
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