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.
Highlights
Nucleic acid-based therapeutics have received significant attention in recent years for the treatment of several diseases, the delivery of therapeutic nucleic acids still remain a major challenge [1]
Double stranded plasmid DNAs can be transfected effectively by using high efficient viral vectors, but, non-viral vectors are generally used for screening single-stranded oligonucleotide in vitro
Despite the pivotal role of non-viral transfection reagents playing in single-stranded oligonucleotide (SSO) screening, a comprehensive comparison of the transfection efficacy of current commercially available transfection reagents has not been reported, they are generally compatible with different types of genetic materials, including antisense oligonucleotide (ASO), siRNA, antimiR, and plasmids
Summary
Nucleic acid-based therapeutics have received significant attention in recent years for the treatment of several diseases, the delivery of therapeutic nucleic acids still remain a major challenge [1]. Double stranded plasmid DNAs can be transfected effectively by using high efficient viral vectors, but, non-viral vectors are generally used for screening single-stranded oligonucleotide in vitro. Despite the pivotal role of non-viral transfection reagents playing in SSO screening, a comprehensive comparison of the transfection efficacy of current commercially available transfection reagents has not been reported, they are generally compatible with different types of genetic materials, including ASO, siRNA, antimiR, and plasmids. The transfection efficiency of non-viral transfection reagents is associated with cell type, cell and media conditions, and. We efficiency report the and transfection comparison of commonly used commercial transfection reagents for in vitro delivery of SSOs ten efficiency and toxicity comparison of commonly used commercial transfection reagents for ininvitro established cell lines, toestablished assist researchers working in this rapidly working progressing fieldrapidly of oligonucleotide delivery of SSOs in ten cell lines, to assist researchers in this progressing therapeutic development.
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