Work

Efficacy of an Amphipathic Oligopeptide to Shuttle and Release a cis -Acting DNA Decoy into Human Cells

Year: 2002

Type: article

Abstract: We investigated the ability of an amphipathic oligopeptide to carry a synthetic dsDNA 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.

Source: BioTechniques

DOI: 10.2144/02321dd03

Authors: L. Citti, P. Rovero, M.G. Colombo, L. Mariani, L. Poliseno, G. Rainaldi

Institutions: National Research Council, University of Salerno, Istituto di Fisiologia Clinica

Language: English

Topic: RNA Interference and Gene Delivery

Subfield: Molecular Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

SDG: Good health and well-being

Open Access status: gold

Estimated APC paid: $1,415