Centre for Comparative Genomics, Murdoch University, Perth, WA 6150, Australia.
Perron Institute for Neurological and Translational Science, Perth, WA 6009, Australia.
Molecules. 2018 Oct 8;23(10):2564. doi: 10.3390/molecules23102564.
Non-viral vector-mediated transfection is a core technique for in vitro screening of oligonucleotides. Despite the growing interests in the development of oliogonucleotide-based drug molecules in recent years, a comprehensive comparison of the transfection efficacy of commonly used commercial transfection reagents has not been reported. In this study, five commonly used transfection reagents, including Lipofectamine 3000, Lipofectamine 2000, Fugene, RNAiMAX and Lipofectin, were comprehensively analyzed in ten cell lines using a fluorescence imaging-based transfection assay. Although the transfection efficacy and toxicity of transfection reagents varied depending on cell types, the toxicity of transfection reagents generally displayed a positive correlation with their transfection efficacy. According to our results, Lipofectamine 3000, Fugene and RNAiMAX showed high transfection efficacy, however, RNAiMAX may be a better option for majority of cells when lower toxicity is desired. The transfection efficacy of Lipofectamine 2000 was compromised by its high toxicity, which may adversely affect its application in most cells. We firmly believe that our findings may contribute to the future In vitro delivery and screening of single-stranded therapeutic oligonucleotides such as antisense oligonucleotides, antimiRs, and DNAzymes.
非病毒载体介导的转染是体外筛选寡核苷酸的核心技术。尽管近年来人们对寡核苷酸类药物分子的开发越来越感兴趣，但尚未有关于常用商业转染试剂转染效率的全面比较。在这项研究中，我们使用基于荧光成像的转染实验，在十种细胞系中综合分析了五种常用的转染试剂，包括 Lipofectamine 3000、Lipofectamine 2000、Fugene、RNAiMAX 和 Lipofectin。尽管转染试剂的转染效率和毒性因细胞类型而异，但转染试剂的毒性通常与其转染效率呈正相关。根据我们的结果，Lipofectamine 3000、Fugene 和 RNAiMAX 显示出较高的转染效率，然而，当需要较低的毒性时，RNAiMAX 可能是大多数细胞的更好选择。Lipofectamine 2000 的转染效率因高毒性而受到影响，这可能会对其在大多数细胞中的应用产生不利影响。我们坚信，我们的研究结果可能有助于未来单链治疗性寡核苷酸（如反义寡核苷酸、抗 miRNA 和 DNA 酶）的体外递送和筛选。
