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
Start Page1674
Date Published04/2018

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.

Short TitleNature Communications