Magnetic Nanoparticles as a Carrier of dsRNA for Gene Therapy

• Published in: Methods in molecular biology (Clifton, N.J.) Vol. 2211; p. 69
• Main Authors: Grabowska, Małgorzata, Grześkowiak, Bartosz F, Rolle, Katarzyna, Mrówczyński, Radosław
• Format: Journal Article
• Language: English
• Published: United States 2021
• Subjects: Cell Line, Tumor; Cell Survival - genetics; Drug Carriers - chemistry; Gene Expression; Gene Transfer Techniques; Genetic Therapy - methods; Humans; Lipids - chemistry; Magnetite Nanoparticles - chemistry; RNA Interference; RNA, Double-Stranded - administration & dosage; RNA, Double-Stranded - genetics; RNA, Small Interfering - administration & dosage; RNA, Small Interfering - genetics; Transfection; RNAi; Transfection; Double-stranded RNA; Glioblastoma; Magnetic nanoparticles; Tenascin-C; Cytotoxicity; Gene therapy
• ISSN: 1940-6029
• DOI: 10.1007/978-1-0716-0943-9_6

Glioma belongs to the most aggressive and lethal types of cancer. Glioblastoma multiforme (GBM), the most common type of malignant gliomas, is characterized by a poor prognosis and remains practically incurable despite aggressive treatment such as surgery, radiotherapy, and chemotherapy. Brain tumor cells overexpress a number of proteins that play a crucial role in tumorigenesis and may be exploited as therapeutic targets. One such target can be an extracellular matrix glycoprotein-tenascin-C (TN-C). Downregulation of TN-C by RNA interference (RNAi) is a very promising strategy in cancer therapy. However, the successful delivery of naked double-stranded RNA (dsRNA) complementary to TN-C sequence (ATN-RNA) requires application of delivery vehicles that can efficiently overcome rapid degradation by nucleases and poor intracellular uptake. Here, we present a protocol for application of MNP@PEI as a carrier for ATN-RNA to GBM cells. The obtained complexes consisted of polyethyleneimine (PEI)-coated magnetic nanoparticles combined with the dsRNA show high efficiency in ATN-RNA delivery, resulting not only in significant TN-C expression level downregulation, but also impairing the tumor cells migration.