Title | RNA-guided transcriptional silencing in vivo with S. aureus CRISPR-Cas9 repressors. |
Publication Type | Journal Article |
Year of Publication | 2018 |
Authors | PI Thakore, JB Kwon, CE Nelson, DC Rouse, MP Gemberling, ML Oliver, and CA Gersbach |
Journal | Nature communications |
Volume | 9 |
Issue | 1 |
Start Page | 1674 |
Date Published | 04/2018 |
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. |
DOI | 10.1038/s41467-018-04048-4 |
Short Title | Nature communications |