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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| Format: | Article |
| Language: | English |
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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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| author | L. Citti P. Rovero M.G. Colombo L. Mariani L. Poliseno G. Rainaldi |
| author_facet | L. Citti P. Rovero M.G. Colombo L. Mariani L. Poliseno G. Rainaldi |
| author_sort | L. Citti |
| collection | DOAJ |
| description | 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. |
| format | Article |
| id | doaj-art-6c6669032b5e4e5b9a26d1d3513273fa |
| institution | OA Journals |
| issn | 0736-6205 1940-9818 |
| language | English |
| publishDate | 2002-01-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | BioTechniques |
| spelling | doaj-art-6c6669032b5e4e5b9a26d1d3513273fa2025-08-20T02:26:07ZengTaylor & Francis GroupBioTechniques0736-62051940-98182002-01-0132117217710.2144/02321dd03Efficacy of an Amphipathic Oligopeptide to Shuttle and Release a cis-Acting DNA Decoy into Human CellsL. Citti0P. Rovero1M.G. Colombo2L. Mariani3L. Poliseno4G. Rainaldi51Istituto di Mutagenesi e Differenziamento, CNR, Pisa, Italy1Istituto di Mutagenesi e Differenziamento, CNR, Pisa, Italy1Istituto di Mutagenesi e Differenziamento, CNR, Pisa, Italy1Istituto di Mutagenesi e Differenziamento, CNR, Pisa, Italy1Istituto di Mutagenesi e Differenziamento, CNR, Pisa, Italy1Istituto di Mutagenesi e Differenziamento, CNR, Pisa, ItalyWe 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.https://www.future-science.com/doi/10.2144/02321dd03 |
| spellingShingle | L. Citti P. Rovero M.G. Colombo L. Mariani L. Poliseno G. Rainaldi Efficacy of an Amphipathic Oligopeptide to Shuttle and Release a cis-Acting DNA Decoy into Human Cells BioTechniques |
| title | Efficacy of an Amphipathic Oligopeptide to Shuttle and Release a cis-Acting DNA Decoy into Human Cells |
| title_full | Efficacy of an Amphipathic Oligopeptide to Shuttle and Release a cis-Acting DNA Decoy into Human Cells |
| title_fullStr | Efficacy of an Amphipathic Oligopeptide to Shuttle and Release a cis-Acting DNA Decoy into Human Cells |
| title_full_unstemmed | Efficacy of an Amphipathic Oligopeptide to Shuttle and Release a cis-Acting DNA Decoy into Human Cells |
| title_short | Efficacy of an Amphipathic Oligopeptide to Shuttle and Release a cis-Acting DNA Decoy into Human Cells |
| title_sort | efficacy of an amphipathic oligopeptide to shuttle and release a cis acting dna decoy into human cells |
| url | https://www.future-science.com/doi/10.2144/02321dd03 |
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