Fluorescence resonance energy transfer analysis of the degradation of an oligonucleotide protected by a very stable hairpin

In vitro degradation of antisense oligonucleotides protected or not on their 3' side against enzymatic attack by a naturally forming hairpin has been studied by fluorescence resonance energy transfer (FRET). The two oligonucleotides d(5"TTCTCGCGAAGC3') forming the hairpin and d(5"...

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Bibliographic Details
Published inJournal of biomolecular structure & dynamics Vol. 14; no. 3; p. 365
Main Authors Réfrégiers, M, Laigle, A, Jollès, B, Chinsky, L
Format Journal Article
LanguageEnglish
Published England 01.12.1996
Subjects
Deoxyribonucleases - metabolism
Electrophoresis, Polyacrylamide Gel
Fluorescein
Fluoresceins - chemistry
Fluorescence
Nucleic Acid Conformation
Oligonucleotides - blood
Oligonucleotides - chemistry
Oligonucleotides - metabolism
Rhodamines - chemistry
Single-Strand Specific DNA and RNA Endonucleases - metabolism
Spectrometry, Fluorescence
Spectrum Analysis - methods
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Summary:In vitro degradation of antisense oligonucleotides protected or not on their 3' side against enzymatic attack by a naturally forming hairpin has been studied by fluorescence resonance energy transfer (FRET). The two oligonucleotides d(5"TTCTCGCGAAGC3') forming the hairpin and d(5"TTCTCCGGAAGC3') as a control were labeled on their 5' side by tetramethylrhodamine and on their 3' side by fluorescein. Fluorescein has been shown not to hinder the hairpin formation and to give an additional protection against nucleases. The FRET technique proved adequate for an in situ study of these protected antisense oligonucleotides in living cells.
ISSN:0739-1102
DOI:10.1080/07391102.1996.10508131