In:
Nature Communications, Springer Science and Business Media LLC, Vol. 9, No. 1 ( 2018-04-26)
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 KRAB ) compatible with adeno-associated viral (AAV) delivery. To evaluate dSaCas9 KRAB 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 KRAB and a Pcsk9 -targeting guide RNA (gRNA) results in significant reductions of serum Pcsk9 and cholesterol levels. Despite a moderate host response to dSaCas9 KRAB 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 KRAB . 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.
Type of Medium:
Online Resource
ISSN:
2041-1723
DOI:
10.1038/s41467-018-04048-4
Language:
English
Publisher:
Springer Science and Business Media LLC
Publication Date:
2018
detail.hit.zdb_id:
2553671-0
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