High-fidelity CRISPR/Cas9- based gene-specific hydroxymethylation rescues gene expression and attenuates renal fibrosis

2018 | journal article; research paper

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​High-fidelity CRISPR/Cas9- based gene-specific hydroxymethylation rescues gene expression and attenuates renal fibrosis​
Xu, X.; Tan, X.; Tampe, B. ; Wilhelmi, T.; Hulshoff, M. S.; Saito, S. & Moser, T.  et al.​ (2018) 
Nature Communications9(1) art. 3509​.​ DOI: https://doi.org/10.1038/s41467-018-05766-5 

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Authors
Xu, Xingbo; Tan, Xiaoying; Tampe, Björn ; Wilhelmi, Tim; Hulshoff, Melanie S.; Saito, Shoji; Moser, Tobias ; Kalluri, Raghu; Hasenfuss, Gerd ; Zeisberg, Elisabeth M. ; Zeisberg, Michael 
Abstract
While suppression of specific genes through aberrant promoter methylation contributes to different diseases including organ fibrosis, gene-specific reactivation technology is not yet available for therapy. TET enzymes catalyze hydroxymethylation of methylated DNA, reactivating gene expression. We here report generation of a high-fidelity CRISPR/Cas9-based gene-specific dioxygenase by fusing an endonuclease deactivated high-fidelity Cas9 (dHFCas9) to TET3 catalytic domain (TET3CD), targeted to specific genes by guiding RNAs (sgRNA). We demonstrate use of this technology in four different anti-fibrotic genes in different cell types in vitro, among them RASAL1 and Klotho, both hypermethylated in kidney fibrosis. Furthermore, in vivo lentiviral delivery of the Rasal1-targeted fusion protein to interstitial cells and of the Klotho-targeted fusion protein to tubular epithelial cells each results in specific gene reactivation and attenuation of fibrosis, providing gene-specific demethylating technology in a disease model.
Issue Date
2018
Journal
Nature Communications 
Project
SFB 1002: Modulatorische Einheiten bei Herzinsuffizienz 
SFB 1002 | C01: Epigenetische Kontrolle der Herzfibrose 
SFB 1002 | D03: ENPP3-vermittelter Phosphat-Metabolismus bei der Herzfibrose 
Working Group
RG Hasenfuß (Transition zur Herzinsuffizienz) 
RG E. Zeisberg (Kardiales Stroma) 
RG M. Zeisberg (Renale Fibrogenese) 
ISSN
2041-1723
Language
English

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