Oligonucleotide-conjugated thiazole orange probes as "light-up" probes for messenger ribonucleic acid molecules in living cells.
"Light-up" probes, icosa-alpha-thymidylate-thiazole orange conjugates, for the in situ time-resolved detection of messenger ribonucleic acid (mRNA) in living cells are evaluated. Upon annealing with polyA in aqueous solutions, the icosa-alpha-thymidylate-thiazole orange conjugates were shown to be up to 15 times more fluorescent. Microinjection of these probes into adherent fibroblasts resulted in high yields of hybridization and fluorescent signals. Incubation of cells in the presence of these probes resulted in facile internalization of the probe and similar painting of the messenger RNA in the nuclear and cytosolic regions.
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Centre de Biophysique Moléculaire, CNRS UPR 4301, Conventionnée avec l'Université d'Orléans et Affilée à l'Inserm, Orléans, France.
Melvin
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Vigny
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D015502:Absorptiometry, Photon; D000818:Animals; D020029:Base Pairing; D052160:Benzothiazoles; D002851:Chromatography, High Pressure Liquid; D001517:Coleoptera / Q000201:enzymology; D015342:DNA Probes / Q000737:chemistry*; D019793:Fluorescein / Q000737:chemistry; D005456:Fluorescent Dyes / Q000032:analysis / Q000737:chemistry; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D008156:Luciferases; D018613:Microscopy, Confocal / Q000379:methods; D008856:Microscopy, Fluorescence; D015394:Molecular Structure; D009693:Nucleic Acid Hybridization / Q000379:methods*; D009841:Oligonucleotides / Q000032:analysis / Q000737:chemistry; D012516:Osteosarcoma; D011804:Quinolines; D012333:RNA, Messenger / Q000032:analysis* / Q000378:metabolism / Q000528:radiation effects; D012343:RNA, Transfer / Q000737:chemistry / Q000378:metabolism / Q000528:radiation effects; D012441:Saccharomyces cerevisiae; D013050:Spectrometry, Fluorescence; D013237:Stereoisomerism; D013844:Thiazoles / Q000737:chemistry; D013997:Time Factors; D014407:Tumor Cells, Cultured / Q000528:radiation effects
D052160:Benzothiazoles; D015342:DNA Probes; D005456:Fluorescent Dyes; D009841:Oligonucleotides; D011804:Quinolines; D012333:RNA, Messenger; D013844:Thiazoles; C051000:thiazole orange; D012343:RNA, Transfer; D008156:Luciferases; D019793:Fluorescein