title: Characterization of oligonucleotide transport into living cells.
abstract: Addition of antisense oligonucleotides to cell cultures has been used to specifically inhibit gene expression. We have investigated the mechanism by which oligonucleotides enter living cells. These compounds are taken up by cells in a saturable, size-dependent manner compatible with receptor-mediated endocytosis. Polynucleotides of any length are competitive inhibitors of oligomer transport, providing they possess a 5'-phosphate moiety. Using oligo(dT)-cellulose for affinity purification, we have identified an 80-kDa surface protein that may mediate transport. Knowledge of the oligonucleotide transport mechanism should facilitate the design of more effective synthetic antisense oligomers as potential clinical agents.
authors.affiliation: Laboratory of Pathology, National Cancer Institute, Bethesda, MD 20892.
authors.forename: S L
authors.initials: SL
authors.lastname: Loke
authors.forename: C A
authors.initials: CA
authors.lastname: Stein
authors.forename: X H
authors.initials: XH
authors.lastname: Zhang
authors.forename: K
authors.initials: K
authors.lastname: Mori
authors.forename: M
authors.initials: M
authors.lastname: Nakanishi
authors.forename: C
authors.initials: C
authors.lastname: Subasinghe
authors.forename: J S
authors.initials: JS
authors.lastname: Cohen
authors.forename: L M
authors.initials: LM
authors.lastname: Neckers
mesh_terms: D000165:Acridine Orange; D001692:Biological Transport / Q000187:drug effects; D002462:Cell Membrane / Q000378:metabolism; D002738:Chloroquine / Q000494:pharmacology; D004705:Endocytosis / Q000187:drug effects; D006801:Humans; D066298:In Vitro Techniques; D007700:Kinetics; D008565:Membrane Proteins / Q000378:metabolism; D008856:Microscopy, Fluorescence; D009841:Oligonucleotides / Q000378:metabolism*; D014407:Tumor Cells, Cultured
chemical_list: D008565:Membrane Proteins; D009841:Oligonucleotides; D002738:Chloroquine; D000165:Acridine Orange