Generation of Cationic Nanoliposomes for the Efficient Delivery of In Vitro Transcribed Messenger RNA
| Autor: | Karlheinz Peter, Stefanie Krajewski, Meike-Kristin Abraham, Antonia Link, Christian Schlensak, Tatjana Michel, Xiaowei Wang, Hans Peter Wendel |
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| Rok vydání: | 2019 |
| Předmět: |
0303 health sciences
Messenger RNA General Immunology and Microbiology Chemistry General Chemical Engineering General Neuroscience Cell RNA Transfection Gene delivery General Biochemistry Genetics and Molecular Biology Green fluorescent protein Cell biology 03 medical and health sciences 0302 clinical medicine medicine.anatomical_structure 030220 oncology & carcinogenesis medicine Protein biosynthesis Viability assay 030304 developmental biology |
| Zdroj: | Journal of Visualized Experiments. |
| ISSN: | 1940-087X |
| DOI: | 10.3791/58444-v |
| Popis: | The development of messenger RNA (mRNA)-based therapeutics for the treatment of various diseases becomes more and more important because of the positive properties of in vitro transcribed (IVT) mRNA. With the help of IVT mRNA, the de novo synthesis of a desired protein can be induced without changing the physiological state of the target cell. Moreover, protein biosynthesis can be precisely controlled due to the transient effect of IVT mRNA. For the efficient transfection of cells, nanoliposomes (NLps) may represent a safe and efficient delivery vehicle for therapeutic mRNA. This study describes a protocol to generate safe and efficient cationic NLps consisting of DC-cholesterol and 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) as a delivery vector for IVT mRNA. NLps having a defined size, a homogeneous distribution, and a high complexation capacity, and can be produced using the dry-film method. Moreover, we present different test systems to analyze their complexation and transfection efficacies using synthetic enhanced green fluorescent protein (eGFP) mRNA, as well as their effect on cell viability. Overall, the presented protocol provides an effective and safe approach for mRNA complexation, which may advance and improve the administration of therapeutic mRNA. |
| Databáze: | OpenAIRE |
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