In Vivo Target Gene Activation via CRISPR/Cas9-Mediated Trans-epigenetic Modulation

• Published in: Cell Vol. 171; no. 7; pp. 1495 - 1507.e15
• Main Authors: Liao, Hsin-Kai, Hatanaka, Fumiyuki, Araoka, Toshikazu, Reddy, Pradeep, Wu, Min-Zu, Sui, Yinghui, Yamauchi, Takayoshi, Sakurai, Masahiro, O’Keefe, David D., Núñez-Delicado, Estrella, Guillen, Pedro, Campistol, Josep M., Wu, Cheng-Jang, Lu, Li-Fan, Esteban, Concepcion Rodriguez, Izpisua Belmonte, Juan Carlos
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 14-12-2017
• Subjects: Animals; Base Sequence; chromatin remodeling; CRISPR-Cas Systems; CRISPR/Cas9; disease model; Disease Models, Animal; Dystrophin - genetics; Epigenesis, Genetic; epigenetic modification; epigenetic therapy; gene editing; Gene Targeting - methods; Genetic Therapy - methods; Interleukin-10 - genetics; Membrane Proteins - genetics; Mice; Mice, Inbred C57BL; muscular dystrophy; Muscular Dystrophy, Duchenne - genetics; Muscular Dystrophy, Duchenne - therapy; regenerative medicine; stem cells; Transcriptional Activation; transdifferentiation; Utrophin - genetics; stem cells; muscular dystrophy; CRISPR/Cas9; regenerative medicine; chromatin remodeling; transdifferentiation; epigenetic modification; gene editing; disease model; epigenetic therapy
• ISSN: 0092-8674; 1097-4172
• DOI: 10.1016/j.cell.2017.10.025

Current genome-editing systems generally rely on inducing DNA double-strand breaks (DSBs). This may limit their utility in clinical therapies, as unwanted mutations caused by DSBs can have deleterious effects. CRISPR/Cas9 system has recently been repurposed to enable target gene activation, allowing regulation of endogenous gene expression without creating DSBs. However, in vivo implementation of this gain-of-function system has proven difficult. Here, we report a robust system for in vivo activation of endogenous target genes through trans-epigenetic remodeling. The system relies on recruitment of Cas9 and transcriptional activation complexes to target loci by modified single guide RNAs. As proof-of-concept, we used this technology to treat mouse models of diabetes, muscular dystrophy, and acute kidney disease. Results demonstrate that CRISPR/Cas9-mediated target gene activation can be achieved in vivo, leading to measurable phenotypes and amelioration of disease symptoms. This establishes new avenues for developing targeted epigenetic therapies against human diseases.
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•A CRISPR/Cas9 system transcriptionally activates endogenous target genes in vivo•Recruiting the transcriptional machinery induces trans-epigenetic remodeling•Inducing target gene expression leads to physiological phenotypes in postnatal mammals•The system ameliorates symptoms associated with several mouse models of human diseases
In vivo delivery of a Cas9-based epigenetic gene activation system ameliorates disease phenotypes in mouse models of type I diabetes, acute kidney injury, and muscular dystrophy