Structure-guided chemical modification of guide RNA enables potent non-viral in vivo genome editing
| Autor: | Victor Koteliansky, Kevin J. Kauffman, Stephen Walsh, Roman L. Bogorad, Hao Yin, Qiongqiong Wu, Alicia Oberholzer, Luke H. Rhym, Daniel G. Anderson, Suet-Yan Kwan, Haiwei Mou, Timofei S. Zatsepin, Robert Langer, Esther Mintzer, Mehmet Fatih Bolukbasi, Sneha Suresh, Lihua Julie Zhu, Wen Xue, Chun-Qing Song, Scot A. Wolfe, Junmei Ding |
|---|---|
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Biomedical Engineering Bioengineering 02 engineering and technology Computational biology Applied Microbiology and Biotechnology Article Mice 03 medical and health sciences Genome editing In vivo Animals Guide RNA Subgenomic mRNA Gene Editing Nuclease Messenger RNA biology Chemistry Cas9 Gene Transfer Techniques RNA 021001 nanoscience & nanotechnology Molecular biology 030104 developmental biology Liver biology.protein Nanoparticles Nucleic Acid Conformation Molecular Medicine CRISPR-Cas Systems Proprotein Convertase 9 0210 nano-technology RNA Guide Kinetoplastida Biotechnology |
| Zdroj: | Nature Biotechnology. 35:1179-1187 |
| ISSN: | 1546-1696 1087-0156 |
| DOI: | 10.1038/nbt.4005 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full text from SpringerLink