論文 - 詳細
| RRC ID | 74389 |
|---|---|
| 著者 | Sakuma S, Okamoto M, Matsushita N, Ukawa M, Tomono T, Kawamoto K, Ikeda T, Sakuma S. |
| タイトル | Evaluation of a D-Octaarginine-linked polymer as a transfection tool for transient and stable transgene expression in human and murine cell lines. |
| ジャーナル | J Vet Med Sci |
| Abstract |
Poly(N-vinylacetamide-co-acrylic acid) coupled with d-octaarginine (VP-R8) promotes the cellular uptake of peptides/proteins in vitro; however, details of the transfection efficacy of VP-R8, such as the cell types possessing high gene transfer, are not known. Herein, we compared the ability of VP-R8 to induce the cellular uptake of plasmid DNA in mouse and human cell lines from different tissues and organs. A green fluorescent protein (GFP)-expression plasmid was used as model genetic material, and fluorescence as an indicator of uptake and plasmid-derived protein expression. Three mouse and three human cell lines were incubated with a mixture of plasmid and VP-R8, and fluorescence analysis were performed two days after transfection. To confirm stable transgene expression, we performed drug selection three days after transfection. A commercially available polymer-based DNA transfection reagent (PTR) was used as the transfection control and standard for comparing transgene expression efficiency. In the case of transient transgene expression, slight-to-moderate GFP expression was observed in all cell lines transfected with plasmid via VP-R8; however, transfection efficiency was lower than using the PTR for gene delivery. In the case of stable transgene expression, VP-R8 promoted drug-resistance acquisition more efficiently than the PTR did. Cells that developed drug resistance after VP-R8-mediated gene transfection expressed GFP more efficiently than cells that developed drug resistance after transfection with the PTR. Thus, VP-R8 shows potential as an in vitro or ex vivo nonviral transfection tool for generating cell lines with stable transgene expression. |
| 巻・号 | 84(4) |
| ページ | 484-493 |
| 公開日 | 2022-4-13 |
| DOI | 10.1292/jvms.21-0647 |
| PMID | 35135938 |
| PMC | PMC9096039 |
| MeSH | Animals Cell Line DNA* Green Fluorescent Proteins / genetics Green Fluorescent Proteins / metabolism Humans Mice Oligopeptides Plasmids / genetics Polymers* Transfection / veterinary Transgenes |
| IF | 1.049 |
| オルトメトリクス指標 |
オルトメトリクス指標項目
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| 各媒体での言及数の合計 | 0 |
| 過去6か月間でのオルトメトリクス指標の変動値 | 0.0 |
| リソース情報 | |
| ヒト・動物細胞 | Hep G2 |