Evaluation of multimeric siRNA conjugates for efficient protamine-based delivery into breast cancer cells

다국어 초록 (Multilingual Abstract)

Despite the preferable properties of well-definedcationic peptides for small interfering RNA (siRNA) delivery,their application as siRNA carriers remains limited dueto their poor binding affinity with short-chain RNAs. In thisstudy, we investigated the feasibility of a novel strategy forcircumventing this limitation, by assessing the utility ofmultimeric conjugates of siRNA for improving the bindingaffinity of siRNAs with cationic peptides and the extent ofintracellular delivery. Protamine, a natural and arginine-richpeptide, was used to produce stably condensed polyelectrolytecomplexes (PECs) with multimeric siRNAs (multisiRNA)with a size of 120 nm while conventional siRNA/protamine particles are over 500 nm. The formulated multisiRNA/protamine PECs showed greatly enhanced stability,intracellular uptake, and biocompatibility compared toconventional, monomeric (mono)-siRNA/protamine particles. With the addition of chloroquine, multi-siRNA/protaminePECs successfully inhibited target gene expression inMDA-MB-435 cells, a breast cancer cell line, even in thepresence of serum protein. This study demonstrates thatmulti-siRNA conjugates greatly facilitate the formulation ofnano-sized protamine-based carriers and significantlyimprove intracellular delivery in vitro compared to commonsiRNAs, and therefore may provide a platform for the designof peptide-based siRNA delivery systems for in vivoapplications.

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