Structure-guided chemical modification of guide RNA enables potent non-viral in vivo genome editing
Name
nihms938717.pdf
Description
Accepted version
Size
1.6 MB
Format
Adobe PDF
Checksum (MD5)
08898800a05ae2290ec52dd9a4d47942
Author(s)
Yin, Hao
Song, Chun-Qing
Suresh, Sneha
Wu, Qiongqiong
Walsh, Stephen C
Rhym, Luke Hyunsik
Mintzer, Esther
Bolukbasi, Mehmet Fatih
Zhu, Lihua Julie
Kauffman, Kevin John
Date Issued
November 2017
Journal
Nature Biotechnology
Publisher
Springer Science and Business Media LLC
Citation
Yin, Hao et al. "Structure-guided chemical modification of guide RNA enables potent non-viral in vivo genome editing." Nature Biotechnology 35, 12 (November 2017): 1179-1187 © 2017 Nature Publishing Group
Version
Author's final manuscript
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.
MIT Department
Massachusetts Institute of Technology. Department of Chemical Engineering
Harvard University--MIT Division of Health Sciences and Technology
Massachusetts Institute of Technology. Institute for Medical Engineering & Science
Massachusetts Institute of Technology. Department of Biology
Koch Institute for Integrative Cancer Research at MIT
Terms of Use
Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use.
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DOI of Published Version
https://doi.org/10.1038/nbt.4005
