Modified oligonucleotides in rabbit reticulocytes: uptake, stability and antisense properties
Identifieur interne : 000509 ( Istex/Corpus ); précédent : 000508; suivant : 000510Modified oligonucleotides in rabbit reticulocytes: uptake, stability and antisense properties
Auteurs : C. Boiziau ; J. J. ToulméSource :
- 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Target region, Thuong, Toulm6, Translation inhibition, Translation initiation, Unmodified, Unmodified oligonucleotides, Unpublished results.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
Links to Exploration step
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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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<ce:abstract-sec><ce:simple-para>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.</ce:simple-para>
</ce:abstract-sec>
</ce:abstract>
<ce:keywords><ce:section-title>Keywords</ce:section-title>
<ce:keyword><ce:text>oligodeoxyribonucleotides</ce:text>
</ce:keyword>
<ce:keyword><ce:text>translation inhibition</ce:text>
</ce:keyword>
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<name type="personal"><namePart type="given">C.</namePart>
<namePart type="family">Boiziau</namePart>
<affiliation>Laboratoire de Biophysique, INSERM U201, Muséum National d'Historie Naturelle, 43, rue Cuvier, 75005 Paris, France</affiliation>
<description>Correspondence and reprints</description>
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<name type="personal"><namePart type="given">J.J.</namePart>
<namePart type="family">Toulmé</namePart>
<affiliation>Laboratoire de Biophysique, INSERM U201, Muséum National d'Historie Naturelle, 43, rue Cuvier, 75005 Paris, France</affiliation>
<description>Correspondence and reprints</description>
<description>Present address: Laboratoire de Biophysique Moléculaire, INSERM CJF 90-13, Université de Bordeaux II, 146 rue Léo-Saignat, F-33076 Bordeaux Cedex, France</description>
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<abstract 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.</abstract>
<subject><genre>Keywords</genre>
<topic>oligodeoxyribonucleotides</topic>
<topic>translation inhibition</topic>
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<dateIssued encoding="w3cdtf">1991</dateIssued>
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<identifier type="ISSN">0300-9084</identifier>
<identifier type="PII">S0300-9084(00)X0152-6</identifier>
<part><date>1991</date>
<detail type="volume"><number>73</number>
<caption>vol.</caption>
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<detail type="issue"><number>11</number>
<caption>no.</caption>
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<extent unit="issue-pages"><start>1339</start>
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