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

Publication ,  Journal Article
Thakore, PI; Kwon, JB; Nelson, CE; Rouse, DC; Gemberling, MP; Oliver, ML; Gersbach, CA
Published in: Nature communications
April 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 (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.

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Published In

Nature communications

DOI

EISSN

2041-1723

ISSN

2041-1723

Publication Date

April 2018

Volume

9

Issue

1

Start / End Page

1674

Related Subject Headings

  • Transcription, Genetic
  • Staphylococcus aureus
  • RNA, Guide, CRISPR-Cas Systems
  • Proprotein Convertase 9
  • Mice, Inbred C57BL
  • Mice
  • Male
  • Liver
  • Genetic Therapy
  • Gene Silencing
 

Citation

APA
Chicago
ICMJE
MLA
NLM
Thakore, P. I., Kwon, J. B., Nelson, C. E., Rouse, D. C., Gemberling, M. P., Oliver, M. L., & Gersbach, C. A. (2018). RNA-guided transcriptional silencing in vivo with S. aureus CRISPR-Cas9 repressors. Nature Communications, 9(1), 1674. https://doi.org/10.1038/s41467-018-04048-4
Thakore, Pratiksha I., Jennifer B. Kwon, Christopher E. Nelson, Douglas C. Rouse, Matthew P. Gemberling, Matthew L. Oliver, and Charles A. Gersbach. “RNA-guided transcriptional silencing in vivo with S. aureus CRISPR-Cas9 repressors.Nature Communications 9, no. 1 (April 2018): 1674. https://doi.org/10.1038/s41467-018-04048-4.
Thakore PI, Kwon JB, Nelson CE, Rouse DC, Gemberling MP, Oliver ML, et al. RNA-guided transcriptional silencing in vivo with S. aureus CRISPR-Cas9 repressors. Nature communications. 2018 Apr;9(1):1674.
Thakore, Pratiksha I., et al. “RNA-guided transcriptional silencing in vivo with S. aureus CRISPR-Cas9 repressors.Nature Communications, vol. 9, no. 1, Apr. 2018, p. 1674. Epmc, doi:10.1038/s41467-018-04048-4.
Thakore PI, Kwon JB, Nelson CE, Rouse DC, Gemberling MP, Oliver ML, Gersbach CA. RNA-guided transcriptional silencing in vivo with S. aureus CRISPR-Cas9 repressors. Nature communications. 2018 Apr;9(1):1674.

Published In

Nature communications

DOI

EISSN

2041-1723

ISSN

2041-1723

Publication Date

April 2018

Volume

9

Issue

1

Start / End Page

1674

Related Subject Headings

  • Transcription, Genetic
  • Staphylococcus aureus
  • RNA, Guide, CRISPR-Cas Systems
  • Proprotein Convertase 9
  • Mice, Inbred C57BL
  • Mice
  • Male
  • Liver
  • Genetic Therapy
  • Gene Silencing