Format

Send to

Choose Destination
See comment in PubMed Commons below
Nat Biotechnol. 2017 Nov 13. doi: 10.1038/nbt.4005. [Epub ahead of print]

Structure-guided chemical modification of guide RNA enables potent non-viral in vivo genome editing.

Author information

1
David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
2
RNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, Massachusetts, USA.
3
Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
4
Department of Molecular, Cell and Cancer Biology, University of Massachusetts Medical School, Worcester, Massachusetts, USA.
5
Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Worcester, Massachusetts, USA.
6
Program in Bioinformatics and Integrative Biology, University of Massachusetts Medical School, Worcester, Massachusetts, USA.
7
Department of Molecular Medicine, University of Massachusetts Medical School, Worcester, Massachusetts, USA.
8
Center of Translational Biomedicine, Skolkovo Institute of Science and Technology, Skolkovo, Moscow, Russia.
9
Department of Chemistry, Lomonosov Moscow State University, Moscow, Russia.
10
Harvard-MIT Division of Health Sciences & Technology, Cambridge, Massachusetts, USA.
11
Institute of Medical Engineering and Science, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.

Abstract

Efficient genome editing with Cas9-sgRNA in vivo has required the use of viral delivery systems, which have limitations for clinical applications. Translational efforts to develop other RNA therapeutics have shown that judicious chemical modification of RNAs can improve therapeutic efficacy by reducing susceptibility to nuclease degradation. Guided by the structure of the Cas9-sgRNA complex, we identify regions of sgRNA that can be modified while maintaining or enhancing genome-editing activity, and we develop an optimal set of chemical modifications for in vivo applications. Using lipid nanoparticle formulations of these enhanced sgRNAs (e-sgRNA) and mRNA encoding Cas9, we show that a single intravenous injection into mice induces >80% editing of Pcsk9 in the liver. Serum Pcsk9 is reduced to undetectable levels, and cholesterol levels are significantly lowered about 35% to 40% in animals. This strategy may enable non-viral, Cas9-based genome editing in the liver in clinical settings.

PMID:
29131148
DOI:
10.1038/nbt.4005
PubMed Commons home

PubMed Commons

0 comments
How to join PubMed Commons

    Supplemental Content

    Full text links

    Icon for Nature Publishing Group
    Loading ...
    Support Center