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10 December 2014 : Original article

New Type of BACE1 siRNA Delivery to Cells

Maciej JabłkowskiAF, Maciej SzemrajACDF, Katarzyna OszajcaCDF, Grażyna JaniszewskaBDF, Jacek BartkowiakABDFG, Janusz SzemrajABCDEFG

DOI: 10.12659/MSM.891219

Med Sci Monit 2014; 20:2598-2606

Abstract

BACKGROUND: Small interfering RNA (siRNA) gene therapy is a new molecular approach in the search for an efficient therapy for Alzheimer disease (AD), based on the principle of RNA interference. Reducing BACE activity can have great therapeutic potential for the treatment of AD. In this study, receptor-mediated delivery was used to deliver opioid peptide-conjugated BACE 1 to INR-32 human neuroblastoma cells.

MATERIAL AND METHODS: An INR-32 human neuroblastoma cell line was stably transfected to express the APP cDNA coding fragment containing the predicted sites for cleavage by α, β, or γ-secretase. This was then treated with BACE 1 siRNA to silence BACE gene expression. BACE gene transcription and translation was determined using BACE-1 siRNA cross-linked with opioid peptide, together with RT-PCR, Western blot analysis, and ELISA.

RESULTS: Receptor-mediated delivery was used to introduce BACE1 siRNA to the APP – INR 32 human neuroblastoma cells. Decreased BACE mRNA and protein expression were observed after the cells were transfected with BACE1 siRNA.

CONCLUSIONS: Delivery of BACE1 siRNA appears to specifically reduce the cleavage of APP by inhibiting BACE1 activity.

Keywords: Amyloid Precursor Protein Secretases - metabolism, Amyloid beta-Protein Precursor - metabolism, Aspartic Acid Endopeptidases - metabolism, Biological Assay, Blotting, Western, Cell Line, Tumor, Gene Expression Regulation, Neoplastic, Gene Transfer Techniques, Genetic Vectors - metabolism, RNA, Messenger - metabolism, RNA, Small Interfering - metabolism, Receptors, Opioid - metabolism, Transfection

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Medical Science Monitor eISSN: 1643-3750
Medical Science Monitor eISSN: 1643-3750