The Neisseria meningitidis CRISPR-Cas9 System Enables Specific Genome Editing in Mammalian Cells

dc.citation.firstpage645
dc.citation.issueNumber3
dc.citation.journalTitleMolecular Therapy
dc.citation.lastpage654
dc.citation.volumeNumber24
dc.contributor.authorLee, Ciaran M.
dc.contributor.authorCradick, Thomas J.
dc.contributor.authorBao, Gang
dc.date.accessioned2017-05-04T18:16:35Z
dc.date.available2017-05-04T18:16:35Z
dc.date.issued2016
dc.description.abstractThe clustered regularly-interspaced short palindromic repeats (CRISPR)—CRISPR-associated (Cas) system from Streptococcus pyogenes (Spy) has been successfully adapted for RNA-guided genome editing in a wide range of organisms. However, numerous reports have indicated that Spy CRISPR-Cas9 systems may have significant off-target cleavage of genomic DNA sequences differing from the intended on-target site. Here, we report the performance of the Neisseria meningitidis (Nme) CRISPR-Cas9 system that requires a longer protospacer-adjacent motif for site-specific cleavage, and present a comparison between the Spy and Nme CRISPR-Cas9 systems targeting the same protospacer sequence. The results with the native crRNA and tracrRNA as well as a chimeric single guide RNA for the Nme CRISPR-Cas9 system were also compared. Our results suggest that, compared with the Spy system, the Nme CRISPR-Cas9 system has similar or lower on-target cleavage activity but a reduced overall off-target effect on a genomic level when sites containing three or fewer mismatches are considered. Thus, the Nme CRISPR-Cas9 system may represent a safer alternative for precision genome engineering applications.
dc.identifier.citationLee, Ciaran M., Cradick, Thomas J. and Bao, Gang. "The Neisseria meningitidis CRISPR-Cas9 System Enables Specific Genome Editing in Mammalian Cells." <i>Molecular Therapy,</i> 24, no. 3 (2016) Elsevier: 645-654. https://doi.org/10.1038/mt.2016.8.
dc.identifier.doihttps://doi.org/10.1038/mt.2016.8
dc.identifier.urihttps://hdl.handle.net/1911/94171
dc.language.isoeng
dc.publisherElsevier
dc.rightsThis work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material.
dc.rights.urihttps://creativecommons.org/licenses/by-nc-sa/4.0/
dc.titleThe Neisseria meningitidis CRISPR-Cas9 System Enables Specific Genome Editing in Mammalian Cells
dc.typeJournal article
dc.type.dcmiText
dc.type.publicationpublisher version
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