Abstract
Messenger RNA (mRNA) has become one of the most potential drugs in recent years. However, efficient and safe delivery of fragile and easily degradable mRNA is a major challenge. Appropriate delivery system (DS) determines the final effect of mRNA. Cationic lipids play a crucial and decisive role in the entire DS, but also cause huge biosafety problems due to the high toxicity. In this study, a new DS for mRNA delivery that combines negatively charged phospholipids was developed in order to neutralize the positive charge and thus increase the safety. Further, the factors affecting mRNA transfection from cell to animal were investigated. The mRNA DS with optimum condition of lipid composition, proportions, structure, and transfection time was synthesized. Adding an appropriate amount of the anionic lipid to liposomes could increase the safety while maintaining the original transfection efficiency. For transporting mRNA in vivo, requirements regarding the mRNA encapsulation and releasing rate should be further considered to optimize DS design and preparation.
Author contributions
Lin Wang, Jun Yang, and Yongan Wang conceived and designed the study. Lin Wang, Huanchun Xing, Shuai Guo, Wenbin Cao, Zinan Zhang, Lijuan Huang and Xin Sui conducted the experiments. Yuan Luo analyzed the data. Jun Yang and Yongan Wang supervised the study and edited the manuscript. All authors read and approved the manuscript.
Disclosure statement
The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.
Data availability
The datasets used or analyzed during the current study are available from the corresponding author on reasonable request.
Ethical approval
All experiments were performed in accordance with the Regulations of the Experimental Animal Administration, issued by the State Committee of Science and Technology of the People’s Republic of China (November 14, 1988), the ARRIVE Guidelines and the Guidelines for Care and Use of Laboratory Animals of the Beijing Institute of Pharmacology and Toxicology. And the experiments were approved by the Animal Ethics Committee of the Beijing Institute of Pharmacology and Toxicology (IACUC permit number: IACUC-DWZX-2023-P649). BALB/c mice (4 weeks old, specific pathogen-free) were acquired from Beijing Charles River Laboratory Animal Technology Co, Ltd. (Beijing, China). Rodents had free access to sterilized food and distilled water and were maintained in stainless steel cages filled with hardwood chips in an air-conditioned room on a 12:12 h light/dark. All procedures were conducted under isoflurane inhalation anesthesia mixed with oxygen while the body temperature was monitored with a feedback-regulated heating pad.