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 Communications, 9(1) art. 3509. DOI: https://doi.org/10.1038/s41467-018-05766-5
Documents & Media
Details
- 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) - External URL
- https://sfb1002.med.uni-goettingen.de/production/literature/publications/225
- ISSN
- 2041-1723
- Language
- English