Intracellular controlled release prolongs the time period of siRNA-based gene suppression

Abstract

RNA interference (RNAi) is a gene silencing process by inhibiting a target messenger RNA (mRNA) in the sequence-specific manner in the cell cytoplasm. Small interfering RNA (siRNA) cleaves the target mRNA. However, siRNA is not generally internalized into cells in the native state. The objective of this study is to prepare cationized gelatin nanospheres (cGNS) incorporating small interfering RNA (siRNA) and to prolong the time period of gene expression suppression. The cGNS with different degradabilities were prepared to evaluate the effect on the suppression of gene expression. There was no difference in the apparent size and zeta potential of cGNS among the amounts of glutaraldehyde (GA) added for crosslinking. The degradation of cGNS tended to become slowly with an increase of GA amounts used in preparation. After MC3T3-E1 cells were incubated with cGNS incorporating siRNA, the gene expression of cells was evaluated by real-time polymerase chain reaction (PCR). The time period of gene suppression increased with an increased amount of siRNA incorporated in cGNS. Moreover, the significant gene suppression was extended over 4 days. It is concluded that the intracellular controlled release with the cGNS enabled siRNA to prolong the time period of gene expression suppression.