A high-throughput screening platform to identify nanocarriers for efficient delivery of RNA-based therapies
| Autor: | Vitor Francisco, Josephine Blersch, Hugo Fernandes, Lino Ferreira, Catarina Rebelo, Artur Filipe Rodrigues |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
Cell type
Polymers media_common.quotation_subject High-throughput screening Cell Computational biology Gene delivery General Biochemistry Genetics and Molecular Biology 03 medical and health sciences Drug Delivery Systems microRNA medicine Internalization Molecular Biology 030304 developmental biology media_common 0303 health sciences Chemistry 030302 biochemistry & molecular biology Gene Transfer Techniques RNA High-Throughput Screening Assays MicroRNAs medicine.anatomical_structure Nanoparticles RNA Long Noncoding Nanocarriers |
| Zdroj: | Methods Repositório Científico de Acesso Aberto de Portugal Repositório Científico de Acesso Aberto de Portugal (RCAAP) instacron:RCAAP CIÊNCIAVITAE |
| ISSN: | 1046-2023 |
| DOI: | 10.1016/j.ymeth.2020.12.002 |
| Popis: | RNA-based therapies are highly selective and powerful regulators of biological functions. Non-viral vectors such as nanoparticles (NPs) are very promising formulations for the delivery of RNA-based therapies but their cell targeting, cell internalization and endolysomal escape capacity is rather limited. Here, we present a methodology that combines high-throughput synthesis of light-triggerable NPs and a high-content imaging screening to identify NPs capable of efficiently delivering different type of RNAs. The NPs were generated using polymers synthesized by Michael type addition reactions and they were designed to: (i) efficiently complex coding (mRNAs) and non-coding (miRNAs and/or lncRNAs) RNA molecules, (ii) allow rapid cell uptake and cytoplasmic release of RNA molecules and (iii) target different cell types based on their composition. Furthermore, light-responsive domains were attached to the polymers by distinctive methods to provide diverse disassembly strategies. The most efficient formulations were identified using cell-based assays and high-content imaging analysis. This strategy allows precise delivery of RNA-based therapies and provides an effective design approach to address critical issues in non-viral gene delivery. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full Text from ScienceDirect