Structure-guided chemical modification of guide RNA enables potent non-viral in vivo genome editing
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; Mou, Haiwei; Oberholzer, Alicia; Ding, Junmei; Kwan, Suet-Yan; Bogorad, Roman; Zatsepin, Timofei; Koteliansky, Victor; Wolfe, Scot A; Xue, Wen; Langer, Robert; Langer, Robert S; Anderson, Daniel Griffith; ... Show more Show less
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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.
DepartmentDavid H. Koch Institute for Integrative Cancer Research at MIT; 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 and Science; Massachusetts Institute of Technology. Department of Biology
Springer Science and Business Media LLC
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
Author's final manuscript