Heavily and Fully Modified RNAs Guide Efficient SpyCas9-Mediated Genome Editing [preprint]
Authors
Mir, AamirAlterman, Julia F.
Hassler, Matthew R.
Debacker, Alexandre J.
Hudgens, Edward
Echeverria, Dimas
Brodsky, Michael H.
Khvorova, Anastasia
Watts, Jonathan K
Sontheimer, Erik J.
UMass Chan Affiliations
Program in Molecular MedicineDepartment of Molecular, Cell and Cancer Biology
RNA Therapeutics Institute
Document Type
PreprintPublication Date
2018-03-28Keywords
RNA-based drugsSpyCas9-Mediated Genome Editing
CRISPR-Cas9-based therapeutics
RNA
crRNA
tracrRNA
biochemistry
Biochemistry
Cell Biology
Genetic Phenomena
Genetics and Genomics
Therapeutics
Metadata
Show full item recordAbstract
RNA-based drugs depend on chemical modifications to increase potency and nuclease stability, 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. No studies have yet explored chemical modification at all positions of the crRNA guide and tracrRNA cofactor. Here, we have identified 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 demonstrate a significant breakthrough for 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.Source
bioRxiv 290999; doi: https://doi.org/10.1101/290999. Link to preprint on bioRxiv service.
DOI
10.1101/290999Permanent Link to this Item
http://hdl.handle.net/20.500.14038/29263Rights
The copyright holder for this preprint (which was not peer-reviewed) is the author/funder. It is made available under a CC-BY 4.0 International license.Distribution License
http://creativecommons.org/licenses/by/4.0/ae974a485f413a2113503eed53cd6c53
10.1101/290999
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Except where otherwise noted, this item's license is described as The copyright holder for this preprint (which was not peer-reviewed) is the author/funder. It is made available under a CC-BY 4.0 International license.