An amphipathic trans-acting phosphorothioate RNA element delivers an uncharged phosphorodiamidate morpholino sequence in mdx mouse myotubes.
作者： H. V. Jain , J. F. Boehler , D. Verthelyi , K. Nagaraju , S. L. Beaucage
DOI: 10.1039/C7RA04247G
关键词: mdx mouse 、 DNA 、 Luciferase 、 Chemistry 、 Transfection 、 Morpholino 、 Myogenesis 、 RNA 、 Exon 、 Molecular biology
摘要: An efficient method for the delivery of uncharged polyA-tailed phosphorodiamidate morpholino sequences (PMO) in mammalian cells consists employing a synthetic 8-mer amphipathic trans-acting poly-2′-O-methyluridylic thiophosphate triester element (2′-OMeUtaPS) as transfection reagent. Unlike dTtaPS DNA-based element, this RNA is potent at delivering PMO to HeLa pLuc 705 or myotube muscle cells. However, much like dTtaPS, 2′-OMeUtaPS-mediated internalization occurs through an energy-dependent mechanism; macropinocytosis appears be predominant endocytic pathway used cellular uptake. The transfected induce alternate splicing either pre-mRNA encoding luciferase excision exon 23 from dystrophin mdx mouse model muscular dystrophy with efficiency comparable that commercial cationic lipid reagents but without detrimental cytotoxicity.
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