https://scholars.lib.ntu.edu.tw/handle/123456789/194419
Title: | 發展肝細胞專一性的蛋白微脂體複合物以作為肝臟基因治療的載體 | Authors: | 倪衍玄 | Keywords: | asialoglycoprotein;protamine;liposome;gene therapy | Issue Date: | 2001 | Publisher: | 臺北市:國立臺灣大學醫學院小兒科 | Abstract: | Background/Aim: Asiaologlycoprotein (AsG) receptors uniquely exist in hepatocytes. Protamine is a polyarginine and able to condense DNA. Liposome is widely used as a gene transfer vector. In this study, we designed a new complex containing AsG-protamine conjugate and mixed with liposome, which uses AsG as a liver specific ligand, protamine to condense DNA, and both protamine and liposome to carry foreign DNA to form an efficient hepatic gene transfer vector. Methods: Conjugation reaction of AsG and protamine and liposome was monitored by capillary electrophoresis. The product then carried green fluorescent protein (GFP) gene to transfect human hepatoma cell line and non-hepatocyte cell line to demonstrate its efficacy and specificity in vitro. For the in vivo study, this gene transfer complex carried GFP gene and injected into the mice either through tail veins or portal veins. The expression of GFP was assayed 48 hours later by the direct fluorescent microscopic examinations and the flow cytometry. Results: AsG-protamine conjugate and liposome mixture had a 2 and 10 more folds increase of GFP expression than liposome or conjugate alone in hepatoma cell line respectively. However, the in vivo studies showed the gene expression level in the liver was minimal either through tail veins or portal veins. Conclusion: AsG-protamine conjugate and liposome mixture is a new liver-specific gene transfer vector. This vector may work well in ex vivo gene therapy and needs further modification to apply to in vivo hepatic gene therapy. |
URI: | http://ntur.lib.ntu.edu.tw//handle/246246/22854 | Other Identifiers: | 892314B002386 | Rights: | 國立臺灣大學醫學院小兒科 |
Appears in Collections: | 醫學系 |
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892314B002386.pdf | 798.14 kB | Adobe PDF | View/Open |
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