TY - JOUR
T1 - Systematic screening of commonly used commercial transfection reagents towards efficient transfection of single-stranded oligonucleotides
AU - Wang, Tao
AU - Larcher, Leon M.
AU - Ma, Lixia
AU - Veedu, Rakesh N.
N1 - Funding Information:
Acknowledgments: R.N.V. greatly acknowledges financial support from the Department of Health (Merit Award) the Western Australian Government; the McCusker Charitable Foundation; and the Perron Institute for Neurological and Translational Science. T.W. is supported by Murdoch University commercial research funding. Authors greatly appreciate and thank Sue Fletcher and Steve Wilton of Murdoch University, Stacey Edwards of QIMR Berghofer Institute, Bruno Meloni of Perron Institute of Neurological and Translational Science, and Willem Lesterhuis of Harry Perkins Institute, University of Western Australia for providing cells as mentioned in the materials and methods section.
Publisher Copyright:
© 2018 MDPI AG. All rights reserved.
PY - 2018/10/8
Y1 - 2018/10/8
N2 - 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.
AB - 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.
KW - Cationic lipid
KW - Cytotoxicity
KW - Gene transfection
KW - single-stranded oligonucleotide
KW - Transfection reagent
UR - http://www.scopus.com/inward/record.url?scp=85054711048&partnerID=8YFLogxK
U2 - 10.3390/molecules23102564
DO - 10.3390/molecules23102564
M3 - Article
C2 - 30297632
SN - 1420-3049
VL - 23
JO - Molecules
JF - Molecules
IS - 10
M1 - 2564
ER -