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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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 A, Alterman JF, Hassler MR, Debacker AJ, Hudgens E, Echeverria D, Brodsky MH, Khvorova A, Watts JK, Sontheimer EJ. (2018). Heavily and fully modified RNAs guide efficient SpyCas9-mediated genome editing. Open Access Publications by UMMS Authors. https://doi.org/10.1038/s41467-018-05073-z. Retrieved from https://escholarship.umassmed.edu/oapubs/3531
Creative Commons License
This work is licensed under a Creative Commons Attribution 4.0 License.