Rationally designed insulator-like elements can block enhancer action in vitro

Insulators are DNA sequences that are likely to be involved in formation of chromatin domains, functional units of gene expression in eukaryotes. Insulators can form domain boundaries and block inappropriate action of regulatory elements (such as transcriptional enhancers) in eukaryotic nuclei. Usin...

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Bibliographic Details
Published in:The EMBO journal 2003-09, Vol.22 (18), p.4728-4737
Main Authors: Bondarenko, Vladimir A., Jiang, Yong I., Studitsky, Vasily M.
Format: Article
Language:English
Subjects:
Base Sequence
Chromatin - genetics
Cloning, Molecular
Deoxyribonucleic acid
DNA
DNA, Superhelical - chemistry
DNA, Superhelical - genetics
Drug Design
EMBO09
Enhancer Elements, Genetic - genetics
enhancers
initiation
Insulation
insulators
Models, Genetic
Molecular Sequence Data
Nucleic Acid Conformation
Promoter Regions, Genetic
Restriction Mapping
RNA polymerase
Templates, Genetic
transcription
Transcription, Genetic
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Summary:Insulators are DNA sequences that are likely to be involved in formation of chromatin domains, functional units of gene expression in eukaryotes. Insulators can form domain boundaries and block inappropriate action of regulatory elements (such as transcriptional enhancers) in eukaryotic nuclei. Using an in vitro system supporting enhancer action over a large distance, the enhancer‐blocking insulator activity has been recapitulated in a highly purified system. The insulator‐like element was constructed using a sequence‐specific DNA‐binding protein making stable DNA loops ( lac repressor). The insulation was entirely dependent on formation of a DNA loop that topologically isolates the enhancer from the promoter. This rationally designed, inducible insulator‐like element recapitulates many key properties of eukaryotic insulators observed in vivo . The data suggest novel mechanisms of enhancer and insulator action.
ISSN:0261-4189
1460-2075
1460-2075
DOI:10.1093/emboj/cdg468