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

BMC Molecular Biology

Nicoletta Corbi¹  Claudio Passananti¹  Elisabetta Mattei²  Lucia Monaco³  Georgios Strimpakos²  Cinzia Pisani¹  Annalisa Onori¹ 

[1] Institute of Molecular Biology and Pathology CNR, c/o Department of Molecular Medicine, University “Sapienza”, Viale Regina Elena 291, 00161, Rome, Italy;Institute of Cell Biology and Neurobiology CNR, IRCCS Fondazione Santa Lucia, Via del Fosso di Fiorano 64, 00143, Rome, Italy;Department of Physiology and Pharmacology, University Sapienza, Piazzale Aldo Moro 5, 00185, Rome, Italy

关键词: Che-1/AATF;    Activation domain;    Artificial transcription factor;    Zinc finger;    Utrophin;    Dystrophin;    DMD;   

Others  :  1091286
DOI  :  10.1186/1471-2199-14-3

 received in 2012-03-30, accepted in 2012-12-12,  发布年份 2013

【 摘 要 】

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.

【 授权许可】

   
2013 Onori et al.; licensee BioMed Central Ltd.

【 图 表 】

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