RESUMO
The potential of RNAi therapies has been largely impeded by the inherent challenges in the functional delivery of siRNA to cells. Herein, we describe protocols for the synthesis and characterization of novel peptide-siRNA nanoparticles prepared from disulfide-constrained amphipathic peptides complexed with siRNA as promising siRNA delivery vectors. We also describe protocols for the application of these nanoparticles to the in vitro and in vivo delivery of siRNA to lung cells for the functional knockdown of lung proteins.
Assuntos
Dissulfetos/química , Sistemas de Liberação de Medicamentos/métodos , Pulmão/efeitos dos fármacos , Nanopartículas/química , Oligonucleotídeos/química , Peptídeos Cíclicos/administração & dosagem , Peptídeos Cíclicos/química , RNA Interferente Pequeno/química , Células A549 , Linhagem Celular Tumoral , Técnicas de Transferência de Genes , Humanos , Interferência de RNA/fisiologiaRESUMO
Hydrogel networks composed of rippled ß-sheet fibrils of coassembled D- and L-Ac-(FKFE)2-NH2 amphipathic peptides exhibit proteolytic stability and increased rheological strength compared to networks of self-assembled L-Ac-(FKFE)2-NH2 pleated ß-sheet fibrils. Modifying the ratios of l and d peptides in the coassembled rippled ß-sheet fibrils alters the degradation profiles of these hydrogel networks.