期刊
JOURNAL OF VETERINARY MEDICAL SCIENCE
卷 84, 期 4, 页码 484-493出版社
JAPAN SOC VET SCI
DOI: 10.1292/jvms.21-0647
关键词
gene-delivery; plasmid-derived protein expression; poly(N-vinylacetamide-co-acrylic acid) bearing D-octaarginine; stable transgene expression; transient transgene expression
资金
- Azabu University
This study compared the ability of VP-R8 to induce cellular uptake of plasmid DNA in mouse and human cell lines from different tissues and organs, showing the potential of VP-R8 as an in vitro or ex vivo nonviral transfection tool.
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.
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