Short, 12 mer fluorescently labeled methylphosphonated oligonucleotides to visualize beta-actin MRNA in vivo.
Identifieur interne : 002300 ( PubMed/Checkpoint ); précédent : 002299; suivant : 002301Short, 12 mer fluorescently labeled methylphosphonated oligonucleotides to visualize beta-actin MRNA in vivo.
Auteurs : K W Lazowski [Pologne] ; L. KaczmarekSource :
- Journal of physiology and pharmacology : an official journal of the Polish Physiological Society [ 0867-5910 ] ; 2003.
Descripteurs français
- KwdFr :
- ADN (), ADN (génétique), ARN messager (analyse), Actines (génétique), Animaux, Cellules 3T3, Colorants fluorescents (), Colorants fluorescents (pharmacologie), Oligodésoxyribonucléotides (), Oligodésoxyribonucléotides (génétique), Organophosphates (), Sondes oligonucléotidiques (normes), Souris, Technique FISH (), Vidéomicroscopie ().
- MESH :
- analyse : ARN messager.
- génétique : ADN, Actines, Oligodésoxyribonucléotides.
- normes : Sondes oligonucléotidiques.
- pharmacologie : Colorants fluorescents.
- ADN, Animaux, Cellules 3T3, Colorants fluorescents, Oligodésoxyribonucléotides, Organophosphates, Souris, Technique FISH, Vidéomicroscopie.
English descriptors
- KwdEn :
- 3T3 Cells, Actins (genetics), Animals, DNA (chemistry), DNA (genetics), Fluorescent Dyes (chemistry), Fluorescent Dyes (pharmacology), In Situ Hybridization, Fluorescence (methods), Mice, Microscopy, Video (methods), Oligodeoxyribonucleotides (chemistry), Oligodeoxyribonucleotides (genetics), Oligonucleotide Probes (standards), Organophosphates (chemistry), RNA, Messenger (analysis).
- MESH :
- chemical , analysis : RNA, Messenger.
- chemical , chemistry : DNA, Fluorescent Dyes, Oligodeoxyribonucleotides, Organophosphates.
- chemical , genetics : Actins, DNA, Oligodeoxyribonucleotides.
- chemical , pharmacology : Fluorescent Dyes.
- methods : In Situ Hybridization, Fluorescence, Microscopy, Video.
- chemical , standards : Oligonucleotide Probes.
- 3T3 Cells, Animals, Mice.
Abstract
A pair of fluorescently labeled antisense ( complementary to beta-actin mRNA) or control methylphosphonated DNA 12-mers were introduced into live cells. After fixation their distribution throughout the cell was compared to the localization pattern for the pair of control oligos.The distribution of the two sets of oligos differed in that there was a distinct pool of antisense probes that were detected at elevated levels in the leading edge of fibroblast and cortical underlining. The resulting fluorescence patterns of antisense probes colocalized and were analogous to labeling pattern already described and produced by in situ hybridization. The length of each of the probe destabilized binding to mismatched sequences at physiological temperature, while the overall length of the pair gave a unique, highly sequence specific recognition of a target sequence. Simultaneous, in vivo application of multiple probes let include internal controls into the experimental setup, in order to distinguish different distributions of antisense and control probes in the same specimen.
PubMed: 14726615
Affiliations:
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pubmed:14726615Le document en format XML
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<author><name sortKey="Lazowski, K W" sort="Lazowski, K W" uniqKey="Lazowski K" first="K W" last="Lazowski">K W Lazowski</name>
<affiliation wicri:level="1"><nlm:affiliation>Laboratory of Molecular Neurobiology, Nencki Institute, Warsaw, Poland. krzych@nencki.gov.pl</nlm:affiliation>
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<author><name sortKey="Kaczmarek, L" sort="Kaczmarek, L" uniqKey="Kaczmarek L" first="L" last="Kaczmarek">L. Kaczmarek</name>
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<term>DNA (chemistry)</term>
<term>DNA (genetics)</term>
<term>Fluorescent Dyes (chemistry)</term>
<term>Fluorescent Dyes (pharmacology)</term>
<term>In Situ Hybridization, Fluorescence (methods)</term>
<term>Mice</term>
<term>Microscopy, Video (methods)</term>
<term>Oligodeoxyribonucleotides (chemistry)</term>
<term>Oligodeoxyribonucleotides (genetics)</term>
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<term>Organophosphates (chemistry)</term>
<term>RNA, Messenger (analysis)</term>
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<term>ADN (génétique)</term>
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<term>Animaux</term>
<term>Cellules 3T3</term>
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<term>Colorants fluorescents (pharmacologie)</term>
<term>Oligodésoxyribonucléotides ()</term>
<term>Oligodésoxyribonucléotides (génétique)</term>
<term>Organophosphates ()</term>
<term>Sondes oligonucléotidiques (normes)</term>
<term>Souris</term>
<term>Technique FISH ()</term>
<term>Vidéomicroscopie ()</term>
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<keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>ADN</term>
<term>Actines</term>
<term>Oligodésoxyribonucléotides</term>
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<term>Cellules 3T3</term>
<term>Colorants fluorescents</term>
<term>Oligodésoxyribonucléotides</term>
<term>Organophosphates</term>
<term>Souris</term>
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<term>Vidéomicroscopie</term>
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<front><div type="abstract" xml:lang="en">A pair of fluorescently labeled antisense ( complementary to beta-actin mRNA) or control methylphosphonated DNA 12-mers were introduced into live cells. After fixation their distribution throughout the cell was compared to the localization pattern for the pair of control oligos.The distribution of the two sets of oligos differed in that there was a distinct pool of antisense probes that were detected at elevated levels in the leading edge of fibroblast and cortical underlining. The resulting fluorescence patterns of antisense probes colocalized and were analogous to labeling pattern already described and produced by in situ hybridization. The length of each of the probe destabilized binding to mismatched sequences at physiological temperature, while the overall length of the pair gave a unique, highly sequence specific recognition of a target sequence. Simultaneous, in vivo application of multiple probes let include internal controls into the experimental setup, in order to distinguish different distributions of antisense and control probes in the same specimen.</div>
</front>
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<Abstract><AbstractText>A pair of fluorescently labeled antisense ( complementary to beta-actin mRNA) or control methylphosphonated DNA 12-mers were introduced into live cells. After fixation their distribution throughout the cell was compared to the localization pattern for the pair of control oligos.The distribution of the two sets of oligos differed in that there was a distinct pool of antisense probes that were detected at elevated levels in the leading edge of fibroblast and cortical underlining. The resulting fluorescence patterns of antisense probes colocalized and were analogous to labeling pattern already described and produced by in situ hybridization. The length of each of the probe destabilized binding to mismatched sequences at physiological temperature, while the overall length of the pair gave a unique, highly sequence specific recognition of a target sequence. Simultaneous, in vivo application of multiple probes let include internal controls into the experimental setup, in order to distinguish different distributions of antisense and control probes in the same specimen.</AbstractText>
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