Document Type
Journal Article
Publication Date
2017
Abstract
An efficient method for the delivery of uncharged polyA-tailed phosphorodiamidate morpholino sequences (PMO) in mammalian cells consists of employing a synthetic 8-mer amphipathic trans-acting poly-2′-O-methyluridylic thiophosphate triester element (2′-OMeUtaPS) as a transfection reagent. Unlike the dTtaPS DNA-based element, this RNA element is potent at delivering polyA-tailed PMO sequences to HeLa pLuc 705 cells or to myotube muscle cells. However, much like dTtaPS, the 2′-OMeUtaPS-mediated internalization of PMO sequences occurs through an energy-dependent mechanism; macropinocytosis appears to be the predominant endocytic pathway used for cellular uptake. The transfected PMO sequences induce alternate splicing of either the pre-mRNA encoding luciferase in HeLa pLuc 705 cells or the excision of exon 23 from the pre-mRNA encoding dystrophin in myotube muscle cells of the mdx mouse model of muscular dystrophy with an efficiency comparable to that of commercial cationic lipid reagents but without detrimental cytotoxicity.
Creative Commons License
This work is licensed under a Creative Commons Attribution-Noncommercial 4.0 License
APA Citation
Jain, H., Boehler, J., Verthelyi, D., Nagaraju, K., & Beaucage, S. (2017). An amphipathic trans-acting phosphorothioate RNA element delivers an uncharged phosphorodiamidate morpholino sequence in mdx mouse myotube. , (). Retrieved from https://hsrc.himmelfarb.gwu.edu/smhs_intsysbio_facpubs/234
Peer Reviewed
1
Open Access
1
Comments
Reproduced with permission of the Royal Society of Chemistry. RSC Advances