Lipofectin-aided cell delivery of ribozyme targeted to human urokinase receptor mRNA.
A 37-mer hammerhead ribozyme has been designed to efficiently cleave the 1.4 kb mRNA of the urokinase plasminogen activator receptor (uPAR). Under in vitro conditions, the chemically synthesized ribozyme cleaved uPAR mRNA and inhibited its translation in a concentration-dependent fashion. The ribozymes were 5'-[35S]thiophosphorylated and used as a model to analyze conditions for RNA delivery in a cultured human osteosarcoma cell system. Ribozymes degraded immediately in cell-conditioned medium but ribozymes complexed with lipofectin were protected from RNases for up to 22 h. Lipofectin rapidly transported ribozyme into the cell, where it accumulated almost exclusively in the cytoplasm. Thus, lipofectin dramatically enhances stability and cytoplasmic delivery of ribozymes, potentially enabling targeting of mRNA in vivo.
Karikó
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Department of Medicine, University of Pennsylvania School of Medicine, Philadelphia 19104-6060.
Megyeri
K
K
Xiao
Q
Q
Barnathan
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ES
D001483:Base Sequence; D002467:Cell Nucleus / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism; D016503:Drug Delivery Systems; D006801:Humans; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D012516:Osteosarcoma / Q000378:metabolism; D010714:Phosphatidylethanolamines; D011131:Polyribonucleotides / Q000138:chemical synthesis; D016337:RNA, Catalytic / Q000737:chemistry / Q000378:metabolism* / Q000493:pharmacokinetics; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism*; D011956:Receptors, Cell Surface / Q000235:genetics*; D055293:Receptors, Urokinase Plasminogen Activator; D014407:Tumor Cells, Cultured
C524898:PLAUR protein, human; D010714:Phosphatidylethanolamines; D011131:Polyribonucleotides; D016337:RNA, Catalytic; D012333:RNA, Messenger; D011956:Receptors, Cell Surface; D055293:Receptors, Urokinase Plasminogen Activator; C020888:1,2-dielaidoylphosphatidylethanolamine