Safe and efficient RNA and DNA introduction into cells using digital electroporation system

Abstract

We systematically compared the delivery and expression efficiencies according to cell types (plant and animal cells) and genetic materials (RNA and DNA) to deliver RNA using a digital electroporation system. Despite the significantly lower RNA delivery in Chlamydomoans reinhartii than DNA delivery due to RNA secondary structure and cell wall, the expression/delivery ratio of RNA was significantly higher than that of DNA (up to 90%), confirming the generally known fact that RNA is more favorable for expression than DNA. On the other hand, in K562 cells, the difference in RNA and DNA delivery efficiency was negligible. Therefore, structural differences between DNA and RNA affect delivery efficiency differently depending on the cell type. RNA delivery efficiency of K562 cells was high, but expression efficiency was much lower than that of microalgae. According to the proposed strategy, compatibility between K562 cells and the nucleic acids used in this study is presumed to be one of the reasons for this low expression efficiency. Gene regulation by delivering small interfering RNA (siRNA) was demonstrated in K562 cells, confirming the feasibility of the digital electroporation system for RNA interference (RNAi) research as a safe and efficient delivery system.

Keywords: Chlamydomoans reinhardtii; Delivery system; Electroporation; K562; RNA delivery.