RNA Therapeutics Institute; Department of Molecular, Cell and Cancer Biology; Program in Molecular Medicine; Department of Biochemistry and Molecular Pharmacology
Biochemistry, Biophysics, and Structural Biology | Genetics and Genomics | Nucleic Acids, Nucleotides, and Nucleosides | Therapeutics
RNA-based drugs depend on chemical modifications to increase potency and to decrease immunogenicity in vivo. Chemical modification will likely improve the guide RNAs involved in CRISPR-Cas9-based therapeutics as well. Cas9 orthologs are RNA-guided microbial effectors that cleave DNA. Here, we explore chemical modifications at all positions of the crRNA guide and tracrRNA cofactor. We identify several heavily modified versions of crRNA and tracrRNA that are more potent than their unmodified counterparts. In addition, we describe fully chemically modified crRNAs and tracrRNAs (containing no 2'-OH groups) that are functional in human cells. These designs will contribute to Cas9-based therapeutics since heavily modified RNAs tend to be more stable in vivo (thus increasing potency). We anticipate that our designs will improve the use of Cas9 via RNP and mRNA delivery for in vivo and ex vivo purposes.
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© The Author(s) 2018. Open Access: This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
DOI of Published Version
Nat Commun. 2018 Jul 6;9(1):2641. doi: 10.1038/s41467-018-05073-z. Link to article on publisher's site
Mir, Aamir; Alterman, Julia F.; Hassler, Matthew R.; Debacker, Alexandre J.; Hudgens, Edward; Echeverria, Dimas; Brodsky, Michael H.; Khvorova, Anastasia; Watts, Jonathan K.; and Sontheimer, Erik J., "Heavily and fully modified RNAs guide efficient SpyCas9-mediated genome editing" (2018). Open Access Articles. 3531.
Creative Commons License
This work is licensed under a Creative Commons Attribution 4.0 License.