Regulation of transcription through light-activation and light-deactivation of triplex-forming oligonucleotides in mammalian cells.
ACS Chem Biol
; 7(7): 1247-56, 2012 Jul 20.
Article
em En
| MEDLINE
| ID: mdl-22540192
ABSTRACT
Triplex-forming oligonucleotides (TFOs) are efficient tools to regulate gene expression through the inhibition of transcription. Here, nucleobase-caging technology was applied to the temporal regulation of transcription through light-activated TFOs. Through site-specific incorporation of caged thymidine nucleotides, the TFODNA triplex formation is blocked, rendering the TFO inactive. However, after a brief UV irradiation, the caging groups are removed, activating the TFO and leading to the inhibition of transcription. Furthermore, the synthesis and site-specific incorporation of caged deoxycytidine nucleotides within TFO inhibitor sequences was developed, allowing for the light-deactivation of TFO function and thus photochemical activation of gene expression. After UV-induced removal of the caging groups, the TFO forms a DNA dumbbell structure, rendering it inactive, releasing it from the DNA, and activating transcription. These are the first examples of light-regulated TFOs and their application in the photochemical activation and deactivation of gene expression. In addition, hairpin loop structures were found to significantly increase the efficacy of phosphodiester DNA-based TFOs in tissue culture.
Texto completo:
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Base de dados:
MEDLINE
Assunto principal:
Oligonucleotídeos
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Estimulação Luminosa
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Transcrição Gênica
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DNA
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Conformação de Ácido Nucleico
Idioma:
En
Ano de publicação:
2012
Tipo de documento:
Article