The use of apolipoprotein A-I as a transport form of the green fluorescent protein GFP gene in rat hepatocytes
The aim of this study was to investigate the possibility of using apolipoprotein A-I (apo A-I) as a transport form of the green fluorescent protein (GFP) gene into rat hepatocytes.Material and methods. A culture of isolated rat hepatocytes was used as a model. Apo A-I conjugate with fluorescein isot...
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Russian Academy of Sciences, Siberian Branch Publishing House
2023-08-01
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| Series: | Сибирский научный медицинский журнал |
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| Online Access: | https://sibmed.elpub.ru/jour/article/view/1164 |
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| author | L. M. Polyakov D. V. Sumenkova M. V. Kotova N. V. Trifonova R. A. Knyazev |
| author_facet | L. M. Polyakov D. V. Sumenkova M. V. Kotova N. V. Trifonova R. A. Knyazev |
| author_sort | L. M. Polyakov |
| collection | DOAJ |
| description | The aim of this study was to investigate the possibility of using apolipoprotein A-I (apo A-I) as a transport form of the green fluorescent protein (GFP) gene into rat hepatocytes.Material and methods. A culture of isolated rat hepatocytes was used as a model. Apo A-I conjugate with fluorescein isothiocyanate (FITC) was obtained by incubation of apo A-I protein with FITC in carbonate buffer pH 9.5 at a ratio of 12.5 μg FITC per 1 mg of protein. Plasmids for pE-GAG transfection with an integrated GFP gene were enriched in the promoter part with cis-elements of the CC(GCC)3-5 type to enhance complex formation with apo A-I. An inverted fluorescence microscope was used for visual analysis of cell fluorescence.Results and discussion. The paper presents evidence of FITC-labeled apo A-I penetration into the cytoplasm and nuclei of rat hepatocytes by receptor-mediated endocytosis. On this basis, it is proposed an attempt to use apo A-I as a means of targeted delivery of plasmid DNA with an integrated GFP gene into the cell. According to the results of fluorescence microscopy, the use of apo A-I as a plasmid DNA transfection agent led to the accumulation of the GFP protein in the cytoplasm of hepatocytes. No fluorescent protein was observed in the absence of apo A-I.Conclusions. The result obtained may indicate the delivery of the GFP gene to the nuclear apparatus of the cell, its expression and GFP protein synthesis. |
| format | Article |
| id | doaj-art-c5af514ec20a45f699f4e07052db881b |
| institution | Kabale University |
| issn | 2410-2512 2410-2520 |
| language | Russian |
| publishDate | 2023-08-01 |
| publisher | Russian Academy of Sciences, Siberian Branch Publishing House |
| record_format | Article |
| series | Сибирский научный медицинский журнал |
| spelling | doaj-art-c5af514ec20a45f699f4e07052db881b2025-08-20T03:50:44ZrusRussian Academy of Sciences, Siberian Branch Publishing HouseСибирский научный медицинский журнал2410-25122410-25202023-08-01434919610.18699/SSMJ20230409521The use of apolipoprotein A-I as a transport form of the green fluorescent protein GFP gene in rat hepatocytesL. M. Polyakov0D. V. Sumenkova1M. V. Kotova2N. V. Trifonova3R. A. Knyazev4Federal Research Center of Fundamental and Translational MedicineNovosibirsk State Medical University of Minzdrav of RussiaFederal Research Center of Fundamental and Translational MedicineFederal Research Center of Fundamental and Translational MedicineFederal Research Center of Fundamental and Translational MedicineThe aim of this study was to investigate the possibility of using apolipoprotein A-I (apo A-I) as a transport form of the green fluorescent protein (GFP) gene into rat hepatocytes.Material and methods. A culture of isolated rat hepatocytes was used as a model. Apo A-I conjugate with fluorescein isothiocyanate (FITC) was obtained by incubation of apo A-I protein with FITC in carbonate buffer pH 9.5 at a ratio of 12.5 μg FITC per 1 mg of protein. Plasmids for pE-GAG transfection with an integrated GFP gene were enriched in the promoter part with cis-elements of the CC(GCC)3-5 type to enhance complex formation with apo A-I. An inverted fluorescence microscope was used for visual analysis of cell fluorescence.Results and discussion. The paper presents evidence of FITC-labeled apo A-I penetration into the cytoplasm and nuclei of rat hepatocytes by receptor-mediated endocytosis. On this basis, it is proposed an attempt to use apo A-I as a means of targeted delivery of plasmid DNA with an integrated GFP gene into the cell. According to the results of fluorescence microscopy, the use of apo A-I as a plasmid DNA transfection agent led to the accumulation of the GFP protein in the cytoplasm of hepatocytes. No fluorescent protein was observed in the absence of apo A-I.Conclusions. The result obtained may indicate the delivery of the GFP gene to the nuclear apparatus of the cell, its expression and GFP protein synthesis.https://sibmed.elpub.ru/jour/article/view/1164apolipoprotein a-iplasmid dnagfp genehepatocytes |
| spellingShingle | L. M. Polyakov D. V. Sumenkova M. V. Kotova N. V. Trifonova R. A. Knyazev The use of apolipoprotein A-I as a transport form of the green fluorescent protein GFP gene in rat hepatocytes Сибирский научный медицинский журнал apolipoprotein a-i plasmid dna gfp gene hepatocytes |
| title | The use of apolipoprotein A-I as a transport form of the green fluorescent protein GFP gene in rat hepatocytes |
| title_full | The use of apolipoprotein A-I as a transport form of the green fluorescent protein GFP gene in rat hepatocytes |
| title_fullStr | The use of apolipoprotein A-I as a transport form of the green fluorescent protein GFP gene in rat hepatocytes |
| title_full_unstemmed | The use of apolipoprotein A-I as a transport form of the green fluorescent protein GFP gene in rat hepatocytes |
| title_short | The use of apolipoprotein A-I as a transport form of the green fluorescent protein GFP gene in rat hepatocytes |
| title_sort | use of apolipoprotein a i as a transport form of the green fluorescent protein gfp gene in rat hepatocytes |
| topic | apolipoprotein a-i plasmid dna gfp gene hepatocytes |
| url | https://sibmed.elpub.ru/jour/article/view/1164 |
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