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

Journal Article (Journal Article)

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 (dSaCas9KRAB ) compatible with adeno-associated viral (AAV) delivery. To evaluate dSaCas9KRAB 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 dSaCas9KRAB and a Pcsk9-targeting guide RNA (gRNA) results in significant reductions of serum Pcsk9 and cholesterol levels. Despite a moderate host response to dSaCas9KRAB 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 dSaCas9KRAB . 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.

Full Text

Duke Authors

Cited Authors

  • Thakore, PI; Kwon, JB; Nelson, CE; Rouse, DC; Gemberling, MP; Oliver, ML; Gersbach, CA

Published Date

  • April 2018

Published In

Volume / Issue

  • 9 / 1

Start / End Page

  • 1674 -

PubMed ID

  • 29700298

Pubmed Central ID

  • PMC5920046

Electronic International Standard Serial Number (EISSN)

  • 2041-1723

International Standard Serial Number (ISSN)

  • 2041-1723

Digital Object Identifier (DOI)

  • 10.1038/s41467-018-04048-4


  • eng