Preparation of Lipid-Conjugated siRNA Oligonucleotides for Enhanced Gene Inhibition in Mammalian Cells

Nucleic acid conjugates are promising drugs for treating gene-related diseases. Conjugating specific units like lipids, cell-penetrating peptides, polymers, antibodies, and aptamers either at the 3′- or 5′-termini of a siRNA duplex molecule has resulted in a plethora of siRNA bioconjugates with impr...

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Detalles Bibliográficos
Autores: Grijalvo, Santiago, Ocampo, Sandra M., Perales, José Carlos, Eritja Casadellà, Ramón
Tipo de recurso: artículo
Fecha de publicación:2021
País:España
Institución:Consejo Superior de Investigaciones Científicas (CSIC)
Repositorio:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/250102
Acceso en línea:http://hdl.handle.net/10261/250102
Access Level:acceso abierto
Palabra clave:Lipids
Ultrasounds
Phosphoramidite chemistry
Serum
RNA interference
siRNA conjugates
siRNA delivery
Descripción
Sumario:Nucleic acid conjugates are promising drugs for treating gene-related diseases. Conjugating specific units like lipids, cell-penetrating peptides, polymers, antibodies, and aptamers either at the 3′- or 5′-termini of a siRNA duplex molecule has resulted in a plethora of siRNA bioconjugates with improved stabilities in bloodstream and better pharmacokinetic values than unmodified siRNAs. In this sense, lipid-siRNA conjugates have attracted a remarkable interest for their potential value in facilitating cellular uptake. In this chapter, we describe a series of protocols involving the synthesis of siRNA oligonucleotides carrying either neutral or cationic lipids at the 3′- and 5′-termini. The resulting lipid-siRNA conjugates are aimed to be used as exogenous effectors for inhibiting gene expression by RNA interference. A protocol for the formulation of lipid siRNA using sonication in the presence of serum is described yielding interesting transfection properties for cell culture without the use of transfecting agents.