UtroUp is a novel six zinc finger artificial transcription factor that recognises 18 base pairs of the utrophin promoter and efficiently drives utrophin upregulation

Annalisa Onori, Cinzia Pisani, Georgios Strimpakos, Lucia Monaco, Elisabetta Mattei, Claudio Passananti, Nicoletta Corbi

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

Background: Duchenne muscular dystrophy (DMD) is the most common X-linked muscle degenerative disease and it is due to the absence of the cytoskeletal protein dystrophin. Currently there is no effective treatment for DMD. Among the different strategies for achieving a functional recovery of the dystrophic muscle, the upregulation of the dystrophin-related gene utrophin is becoming more and more feasible.Results: We have previously shown that the zinc finger-based artificial transcriptional factor " Jazz" corrects the dystrophic pathology in mdx mice by upregulating utrophin gene expression. Here we describe a novel artificial transcription factor, named " UtroUp" , engineered to further improve the DNA-binding specificity. UtroUp has been designed to recognise an extended DNA target sequence on both the human and mouse utrophin gene promoters. The UtroUp DNA-binding domain contains six zinc finger motifs in tandem, which is able to recognise an 18-base-pair DNA target sequence that statistically is present only once in the human genome. To achieve a higher transcriptional activation, we coupled the UtroUp DNA-binding domain with the innovative transcriptional activation domain, which was derived from the multivalent adaptor protein Che-1/AATF. We show that the artificial transcription factor UtroUp, due to its six zinc finger tandem motif, possesses a low dissociation constant that is consistent with a strong affinity/specificity toward its DNA-binding site. When expressed in mammalian cell lines, UtroUp promotes utrophin transcription and efficiently accesses active chromatin promoting accumulation of the acetylated form of histone H3 in the utrophin promoter locus.Conclusions: This novel artificial molecule may represent an improved platform for the development of future applications in DMD treatment.

Original languageEnglish
Article number3
JournalBMC Molecular Biology
Volume14
DOIs
Publication statusPublished - Jan 30 2013

Fingerprint

Utrophin
Zinc Fingers
Base Pairing
Duchenne Muscular Dystrophy
Transcription Factors
Up-Regulation
Dystrophin
DNA
Transcriptional Activation
Inbred mdx Mouse
Muscles
Cytoskeletal Proteins
Human Genome
Histones
Genes
Chromatin
Binding Sites
Pathology
Gene Expression
Cell Line

Keywords

  • Activation domain
  • Artificial transcription factor
  • Che-1/AATF
  • DMD
  • Dystrophin
  • Utrophin
  • Zinc finger

ASJC Scopus subject areas

  • Molecular Biology

Cite this

UtroUp is a novel six zinc finger artificial transcription factor that recognises 18 base pairs of the utrophin promoter and efficiently drives utrophin upregulation. / Onori, Annalisa; Pisani, Cinzia; Strimpakos, Georgios; Monaco, Lucia; Mattei, Elisabetta; Passananti, Claudio; Corbi, Nicoletta.

In: BMC Molecular Biology, Vol. 14, 3, 30.01.2013.

Research output: Contribution to journalArticle

Onori, Annalisa ; Pisani, Cinzia ; Strimpakos, Georgios ; Monaco, Lucia ; Mattei, Elisabetta ; Passananti, Claudio ; Corbi, Nicoletta. / UtroUp is a novel six zinc finger artificial transcription factor that recognises 18 base pairs of the utrophin promoter and efficiently drives utrophin upregulation. In: BMC Molecular Biology. 2013 ; Vol. 14.
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AU - Onori, Annalisa

AU - Pisani, Cinzia

AU - Strimpakos, Georgios

AU - Monaco, Lucia

AU - Mattei, Elisabetta

AU - Passananti, Claudio

AU - Corbi, Nicoletta

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N2 - Background: Duchenne muscular dystrophy (DMD) is the most common X-linked muscle degenerative disease and it is due to the absence of the cytoskeletal protein dystrophin. Currently there is no effective treatment for DMD. Among the different strategies for achieving a functional recovery of the dystrophic muscle, the upregulation of the dystrophin-related gene utrophin is becoming more and more feasible.Results: We have previously shown that the zinc finger-based artificial transcriptional factor " Jazz" corrects the dystrophic pathology in mdx mice by upregulating utrophin gene expression. Here we describe a novel artificial transcription factor, named " UtroUp" , engineered to further improve the DNA-binding specificity. UtroUp has been designed to recognise an extended DNA target sequence on both the human and mouse utrophin gene promoters. The UtroUp DNA-binding domain contains six zinc finger motifs in tandem, which is able to recognise an 18-base-pair DNA target sequence that statistically is present only once in the human genome. To achieve a higher transcriptional activation, we coupled the UtroUp DNA-binding domain with the innovative transcriptional activation domain, which was derived from the multivalent adaptor protein Che-1/AATF. We show that the artificial transcription factor UtroUp, due to its six zinc finger tandem motif, possesses a low dissociation constant that is consistent with a strong affinity/specificity toward its DNA-binding site. When expressed in mammalian cell lines, UtroUp promotes utrophin transcription and efficiently accesses active chromatin promoting accumulation of the acetylated form of histone H3 in the utrophin promoter locus.Conclusions: This novel artificial molecule may represent an improved platform for the development of future applications in DMD treatment.

AB - Background: Duchenne muscular dystrophy (DMD) is the most common X-linked muscle degenerative disease and it is due to the absence of the cytoskeletal protein dystrophin. Currently there is no effective treatment for DMD. Among the different strategies for achieving a functional recovery of the dystrophic muscle, the upregulation of the dystrophin-related gene utrophin is becoming more and more feasible.Results: We have previously shown that the zinc finger-based artificial transcriptional factor " Jazz" corrects the dystrophic pathology in mdx mice by upregulating utrophin gene expression. Here we describe a novel artificial transcription factor, named " UtroUp" , engineered to further improve the DNA-binding specificity. UtroUp has been designed to recognise an extended DNA target sequence on both the human and mouse utrophin gene promoters. The UtroUp DNA-binding domain contains six zinc finger motifs in tandem, which is able to recognise an 18-base-pair DNA target sequence that statistically is present only once in the human genome. To achieve a higher transcriptional activation, we coupled the UtroUp DNA-binding domain with the innovative transcriptional activation domain, which was derived from the multivalent adaptor protein Che-1/AATF. We show that the artificial transcription factor UtroUp, due to its six zinc finger tandem motif, possesses a low dissociation constant that is consistent with a strong affinity/specificity toward its DNA-binding site. When expressed in mammalian cell lines, UtroUp promotes utrophin transcription and efficiently accesses active chromatin promoting accumulation of the acetylated form of histone H3 in the utrophin promoter locus.Conclusions: This novel artificial molecule may represent an improved platform for the development of future applications in DMD treatment.

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