Abstract
Chitosan and its derivatives have been extensively utilized in gene delivery applications because of their low toxicity and positively charged characteristics. However, their low solubility under physiological conditions often limits their application. Glycol chitosan (GC) is a derivative of chitosan that exhibits excellent solubility in physiological buffer solutions. However, it lacks the positive characteristics of a gene carrier. Thus, we hypothesized that the introduction of oligoarginine peptide to GC could improve the formation of complexes with siRNA, resulting in enhanced uptake by cells and increased transfection efficiency in vitro. A peptide with nine arginine residues and 10 glycine units (R9G10) was successfully conjugated to GC, which was confirmed by infrared spectroscopy, 1H NMR spectroscopy, and elemental analysis. The physicochemical characteristics of R9G10-GC/siRNA complexes were also investigated. The size and surface charge of the R9G10-GC/siRNA nanoparticles depended on the amount of R9G10 coupled to the GC. In addition, the R9G10-GC/siRNA nanoparticles showed improved uptake in HeLa cells and enhanced in vitro transfection efficiency while maintaining low cytotoxicity determined by the MTT assay. Oligoarginine-modified glycol chitosan may be useful as a potential gene carrier in many therapeutic applications.
Highlights
Accepted: 30 November 2021RNA interference (RNAi) is an innovative approach for selective and specific gene silencing and has demonstrated potential for the treatment of various diseases [1,2,3,4,5]
ethyl-3-(dimethylaminopropyl) carbodiimide (EDC) is a representative zero-length cross-linker and its conjugation reaction can be improved in the presence of sulfo-NHS because of the prolonged stability of the active ester intermediate against hydrolysis in aqueous solution when compared with the EDC [41]
We demonstrated that nonaarginine-modified glycol chitosan was useful for small interfering RNA (siRNA)
Summary
RNA interference (RNAi) is an innovative approach for selective and specific gene silencing and has demonstrated potential for the treatment of various diseases [1,2,3,4,5]. In this pathway, RNA-induced silencing complex (RISC), formed from double-stranded small interfering RNA (siRNA), degrades target mRNA and inhibits protein synthesis [6,7]. RNAi has often been used to treat many disorders, including cancer, infectious diseases, and rare genetic disorders [8,9,10,11]. Many siRNA-based nanotherapeutics have been investigated clinically to treat patients and several of them are approved by FDA (e.g., ONPATTRO® , Alnylam, Cambridge, MA, USA) [14,15,16]
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