title: The Neisseria meningitidis CRISPR-Cas9 System Enables Specific Genome Editing in Mammalian Cells.
abstract: The 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.
authors.affiliation: Department of Bioengineering, Rice University, Houston, Texas, USA.
authors.forename: Ciaran M
authors.initials: CM
authors.lastname: Lee
authors.affiliation: Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, Georgia, USA.|Present Address: CRISPR Therapeutics, Cambridge, Massachusetts, USA.
authors.forename: Thomas J
authors.initials: TJ
authors.lastname: Cradick
authors.affiliation: Department of Bioengineering, Rice University, Houston, Texas, USA.
authors.forename: Gang
authors.initials: G
authors.lastname: Bao
mesh_terms: D001483:Base Sequence; D001665:Binding Sites; D064113:CRISPR-Cas Systems; D064112:Clustered Regularly Interspaced Short Palindromic Repeats; D000072669:Gene Editing; D018390:Gene Targeting; D056426:Genetic Loci; D016678:Genome; D006801:Humans; D009345:Neisseria meningitidis / Q000201:enzymology* / Q000235:genetics*; D059372:Nucleotide Motifs; D056510:Position-Specific Scoring Matrices; D011485:Protein Binding; D000094704:RNA, Guide, CRISPR-Cas Systems; D013379:Substrate Specificity
chemical_list: D000094704:RNA, Guide, CRISPR-Cas Systems