Modified oligonucleotides in rabbit reticulocytes: uptake, stability and antisense properties
Identifieur interne : 004920 ( Main/Exploration ); précédent : 004919; suivant : 004921Modified oligonucleotides in rabbit reticulocytes: uptake, stability and antisense properties
Auteurs : C. Boiziau [France] ; J. J. Toulmé [France]Source :
- Biochimie [ 0300-9084 ] ; 1991.
English descriptors
- Teeft :
- Acad, Analogue, Antisense, Antisense oligonucleotide, Antisense oligonucleotides, Antisense properties, Biol chem, Blood cells, Coding region, Coding sequence, Comparative inhibition, Culture medium, Eagle medium, External concentration, Frog oocytes, Gene expression, Globin, Globin mrna, Globin synthesis, Growth medium, Independent mechanism, Initiation codon, Intact reticulocytes, Intercalating agents, Intracellular concentration, Large part, Mrna, Natl, Nucleic acids, Oligomer, Oligomers, Oligonucleotide, Oligonucleotides, Phosphorothioate, Phosphorothioate analogues, Phosphorothioate derivatives, Proc natl acad, Protein synthesis, Rabbit globin mrna translation, Rabbit mrna, Rabbit reticulocyte lysate, Rabbit reticulocytes, Reticulocyte, Reticulocyte lysate, Secondary structures, Specific inhibition, Target location, Target region, Thuong, Toulm6, Translation inhibition, Translation initiation, Unmodified, Unmodified oligonucleotides, Unpublished results.
Abstract
Abstract: We have investigated the behaviour of antisense oligonucleotides in rabbit reticulocytes. Both backbone-modified oligomers (methyl-phosphonate and phosphorothio ate analogues), anomers of nucleotide units (α) and oligonucleotides linked to various ligands (intercalator, polylysine, dodecanol) were tested with respect to cellular uptake and inhibition of protein synthesis. Oligonucleotides added at an external concentration of 10 μM slowly entered the cell up to a concentration of a few hundred nanomolars. A large fraction of phosphorothioate analogues was found to be associated with the membrane. α-, methylphosphonate and phosphorothioate analogues remained intact during the incubation with reticulocytes although the latter were partly dephosphorylated. Antisense oligonucleotides were targeted against three different sites of the rabbit β-globin mRNA: the 5′ end of the message, the initiator AUG or the coding sequence. No specific effect on β-globin synthesis was observed with any of the investigated compounds.
Url:
DOI: 10.1016/0300-9084(91)90171-V
Affiliations:
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Le document en format XML
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<front><div type="abstract" xml:lang="en">Abstract: We have investigated the behaviour of antisense oligonucleotides in rabbit reticulocytes. Both backbone-modified oligomers (methyl-phosphonate and phosphorothio ate analogues), anomers of nucleotide units (α) and oligonucleotides linked to various ligands (intercalator, polylysine, dodecanol) were tested with respect to cellular uptake and inhibition of protein synthesis. Oligonucleotides added at an external concentration of 10 μM slowly entered the cell up to a concentration of a few hundred nanomolars. A large fraction of phosphorothioate analogues was found to be associated with the membrane. α-, methylphosphonate and phosphorothioate analogues remained intact during the incubation with reticulocytes although the latter were partly dephosphorylated. Antisense oligonucleotides were targeted against three different sites of the rabbit β-globin mRNA: the 5′ end of the message, the initiator AUG or the coding sequence. No specific effect on β-globin synthesis was observed with any of the investigated compounds.</div>
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