Efficacy of an Amphipathic Oligopeptide to Shuttle and Release a cis-Acting DNA Decoy into Human Cells
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...
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| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Taylor & Francis Group
2002-01-01
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| Series: | BioTechniques |
| Online Access: | https://www.future-science.com/doi/10.2144/02321dd03 |
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| Summary: | 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. |
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| ISSN: | 0736-6205 1940-9818 |