Efficacy of an Amphipathic Oligopeptide to Shuttle and Release a cis-Acting DNA Decoy into Human Cells
| Autor: | L. Citti, P. Rovero, M.G. Colombo, L. Mariani, L. Poliseno, G. Rainaldi |
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| Jazyk: | angličtina |
| Rok vydání: | 2002 |
| Předmět: | |
| Zdroj: | BioTechniques, Vol 32, Iss 1, Pp 172-177 (2002) |
| Druh dokumentu: | article |
| ISSN: | 1940-9818 0736-6205 |
| DOI: | 10.2144/02321dd03 |
| Popis: | We investigated the ability of an amphipathic oligopeptide to carry a synthetic ds- DNA oligonucleotide inside human cells. The oligonucleotide was designed as a decoy binding site for the transcriptional activator of the methylguanine-DNA methyltransferase (MGMT) gene. The complex oligopeptide and decoy were administered to MCF10A exponentially growing cells, and the uptake was monitored by flow cytometry. After a 1-h exposure, almost all of the MCF10A cells were fluorescent, indicating that all of the cells had been transfected. By increasing the time, the fluorescence intensity per cell rapidly increased to a plateau at the 8-h time point. RT-PCR analysis of the MGMT gene was used as the molecular readout of the intracellular activity of the DNA decoy. MCF10A cells transfected with the oligopeptide/decoy complex showed a strong reduction in MGMT mRNA. Here, we discuss the advantages of using amphipathic oligopeptides as carriers of short DNA sequences. |
| Databáze: | Directory of Open Access Journals |
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