Efficacy of an Amphipathic Oligopeptide to Shuttle and Release a cis-Acting DNA Decoy into Human Cells. (January 2002)

• Record Type: Journal Article
• Authors: Citti, L.; Rovero, P.; Colombo, M.G.; Mariani, L.; Poliseno, L.; Rainaldi, G.
• Is Part Of: Biotechniques. Volume 32:Number 1(2002)
• Page Start: 172
• Page End: 177
• Publication Date: 2002-01
• Subjects: Biology, Experimental -- Periodicals; Molecular biology -- Periodicals; Medical technology -- Periodicals; Biology, Experimental; Medical technology; Molecular biology; Clinical Laboratory Techniques -- Periodicals; Research -- Periodicals; Medical Laboratory Science -- Periodicals; Periodicals; Electronic journals; 570
• DOI: 10.2144/02321dd03
• Languages: English
• ISSNs: 0736-6205

Abstract

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.