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Molecules 2018, 23(10), 2564; https://doi.org/10.3390/molecules23102564

Systematic Screening of Commonly Used Commercial Transfection Reagents towards Efficient Transfection of Single-Stranded Oligonucleotides

1,2,†, 1,†, 3 and 1,2,*
1
Centre for Comparative Genomics, Murdoch University, Perth, WA 6150, Australia
2
Perron Institute for Neurological and Translational Science, Perth, WA 6009, Australia
3
School of Statistics, Henan University of Economics and Law, Zhengzhou 450046, China
*
Author to whom correspondence should be addressed.
These authors contributed equally to this paper.
Received: 10 September 2018 / Revised: 4 October 2018 / Accepted: 5 October 2018 / Published: 8 October 2018
(This article belongs to the Special Issue Antisense Oligonucleotide Chemistry and Applications)
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Abstract

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. View Full-Text
Keywords: single-stranded oligonucleotide; transfection reagent; cationic lipid; gene transfection; cytotoxicity single-stranded oligonucleotide; transfection reagent; cationic lipid; gene transfection; cytotoxicity
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Wang, T.; Larcher, L.M.; Ma, L.; Veedu, R.N. Systematic Screening of Commonly Used Commercial Transfection Reagents towards Efficient Transfection of Single-Stranded Oligonucleotides. Molecules 2018, 23, 2564.

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