RNA-guided transcriptional silencing in vivo with S. aureus CRISPR-Cas9 repressors.

TitleRNA-guided transcriptional silencing in vivo with S. aureus CRISPR-Cas9 repressors.
Publication TypeJournal Article
Year of Publication2018
AuthorsPI Thakore, JB Kwon, CE Nelson, DC Rouse, MP Gemberling, ML Oliver, and CA Gersbach
JournalNature Communications
Volume9
Start Page1674
Issue1
Date Published04/2018
Abstract

CRISPR-Cas9 transcriptional repressors have emerged as robust tools for disrupting gene regulation in vitro but have not yet been adapted for systemic delivery in adult animal models. Here we describe a Staphylococcus aureus Cas9-based repressor (dSaCas9<sup>KRAB</sup>) compatible with adeno-associated viral (AAV) delivery. To evaluate dSaCas9<sup>KRAB</sup> efficacy for gene silencing in vivo, we silenced transcription of Pcsk9, a regulator of cholesterol levels, in the liver of adult mice. Systemic administration of a dual-vector AAV8 system expressing dSaCas9<sup>KRAB</sup> and a Pcsk9-targeting guide RNA (gRNA) results in significant reductions of serum Pcsk9 and cholesterol levels. Despite a moderate host response to dSaCas9<sup>KRAB</sup> expression, Pcsk9 repression is maintained for 24 weeks after a single treatment, demonstrating the potential for long-term gene silencing in post-mitotic tissues with dSaCas9<sup>KRAB</sup>. In vivo programmable gene silencing enables studies that link gene regulation to complex phenotypes and expands the CRISPR-Cas9 perturbation toolbox for basic research and gene therapy applications.

DOI10.1038/s41467-018-04048-4
Short TitleNature Communications