Structure-guided chemical modification of guide RNA enables potent non-viral in vivo genome editing
Bolukbasi, Mehmet F.
Zhu, Lihua Julie
Wolfe, Scot A.
Anderson, Daniel G.
UMass Chan AffiliationsProgram in Molecular Medicine
Department of Biochemistry and Molecular Pharmacology
Department of Molecular, Cell and Cancer Biology
RNA Therapeutics Institute
Document TypeJournal Article
KeywordsBiochemistry, Biophysics, and Structural Biology
Cell and Developmental Biology
Genetics and Genomics
MetadataShow full item record
AbstractEfficient 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.
Nat Biotechnol. 2017 Dec;35(12):1179-1187. doi: 10.1038/nbt.4005. Epub 2017 Nov 13. Link to article on publisher's site
Permanent Link to this Itemhttp://hdl.handle.net/20.500.14038/48851
Full list of authors omitted for brevity. For full list see article.