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

Molecular Therapy : the Journal of the American Society of Gene Therapy
Ciaran M LeeGang Bao

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.

References

Mar 24, 2007·Science·Rodolphe BarrangouPhilippe Horvath
Jul 28, 2010·Genetics·Michelle ChristianDaniel F Voytas
Apr 10, 2012·Nature Biotechnology·Deepak ReyonJ Keith Joung
Jun 30, 2012·Science·Martin JinekEmmanuelle Charpentier
Jan 5, 2013·Science·Le CongFeng Zhang
Jan 5, 2013·Science·Prashant MaliGeorge M Church
Jan 31, 2013·Nature Biotechnology·Woong Y HwangJ Keith Joung
Jan 31, 2013·Nature Biotechnology·Wenyan JiangLuciano A Marraffini
Jan 31, 2013·Nature Biotechnology·Seung Woo ChoJin-Soo Kim
Mar 6, 2013·Nucleic Acids Research·James E DiCarloGeorge M Church
Apr 3, 2013·Cell Research·Bin ShenXingxu Huang
Jun 25, 2013·Nature Biotechnology·Yanfang FuJeffry D Sander
Jul 3, 2013·Nature Methods·Ari E FriedlandJohn A Calarco
Jul 23, 2013·Nature Biotechnology·Patrick D HsuFeng Zhang
Aug 10, 2013·Nature Biotechnology·Vladimir NekrasovSophien Kamoun
Aug 14, 2013·Nucleic Acids Research·Thomas J CradickGang Bao
Aug 14, 2013·Proceedings of the National Academy of Sciences of the United States of America·Zhonggang HouJames A Thomson
Oct 1, 2013·Nature Methods·Kevin M EsveltGeorge M Church
Jan 28, 2014·Nature Biotechnology·Yanfang FuJ Keith Joung
Feb 18, 2014·Cell·Hiroshi NishimasuOsamu Nureki
Apr 23, 2014·Nature Biotechnology·Xuebing WuPhillip A Sharp
Apr 29, 2014·Nature Biotechnology·Shengdar Q TsaiJ Keith Joung
Jun 28, 2014·Nature Methods·Sangsu BaeJin-Soo Kim
Dec 3, 2014·Molecular Therapy. Nucleic Acids·Thomas J CradickGang Bao
Dec 17, 2014·Nature Biotechnology·Richard L FrockFrederick W Alt
Dec 17, 2014·Nature Biotechnology·Shengdar Q TsaiJ Keith Joung
Apr 2, 2015·Nature·F Ann RanFeng Zhang
Sep 1, 2015·Cell·Hiroshi NishimasuOsamu Nureki
Oct 17, 2015·Molecular Cell·Yan ZhangErik J Sontheimer
Nov 3, 2015·Nature Biotechnology·Benjamin P KleinstiverJ Keith Joung
Nov 7, 2015·Molecular Cell·Enbo MaJennifer A Doudna

❮ Previous
Next ❯

Citations

May 4, 2016·Plant Cell Reports·Scott M Schaeffer, Paul A Nakata
Jun 3, 2016·Frontiers in Plant Science·Yuduan DingKabin Xie
Jan 12, 2016·Molecular Therapy : the Journal of the American Society of Gene Therapy·Ciaran M LeeGang Bao
Oct 14, 2016·Nucleic Acids Research·Chance M NowakLeonidas Bleris
Jan 11, 2017·Hämostaseologie·Simone A HaasToni Cathomen
Apr 8, 2017·Nature Medicine·Tatjana I CornuToni Cathomen
Apr 4, 2017·Pathogens and Disease·Yan Zhang
Apr 5, 2017·Annual Review of Biophysics·Fuguo Jiang, Jennifer A Doudna
Mar 17, 2017·Scientific Reports·Kelsey E JarrettWilliam R Lagor
Jan 24, 2019·Cellular and Molecular Life Sciences : CMLS·Dmitry KostyushevVladimir Chulanov
Apr 16, 2019·Nucleic Acids Research·Mareike D HoffmannDominik Niopek
Apr 24, 2019·Journal of Neuroimmune Pharmacology : the Official Journal of the Society on NeuroImmune Pharmacology·Sudhanshu P RaikwarAsgar Zaheer
May 12, 2018·Laboratory Investigation; a Journal of Technical Methods and Pathology·Alvin Ho-Kwan CheungKa-Fai To
May 28, 2019·Molecular Genetics and Genomics : MGG·Jinzhao LongGuangcai Duan
Jul 17, 2019·Nature Plants·Yingxiao ZhangYiping Qi
Sep 8, 2019·Nature Communications·Natalia Gomez-OspinaMatthew H Porteus
Feb 7, 2020·Archives of Insect Biochemistry and Physiology·Xu-Yang NiXiaomu Qiao
Feb 11, 2020·Journal of Cellular and Molecular Medicine·Jingyi WangBo Feng
Jun 21, 2019·Journal of Cellular Biochemistry·Vahideh AhmadzadehHabib Zarredar
Oct 28, 2019·TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik·Ke WangXingguo Ye
Jul 10, 2020·The Plant Journal : for Cell and Molecular Biology·Jia DongMatthew E Hudson
Sep 30, 2020·RNA Biology·Yadan Zhang, Mario Andrea Marchisio
Apr 4, 2019·Molecular Biology Reports·Rakesh KumarG P Singh
Jan 17, 2020·Nucleic Acids Research·Iana FedorovaKonstantin Severinov
Jun 12, 2017·European Journal of Clinical Microbiology & Infectious Diseases : Official Publication of the European Society of Clinical Microbiology·P GholizadehH S Kafil
Aug 2, 2018·Frontiers in Plant Science·Deepa JaganathanGayatri Venkataraman
Sep 3, 2020·International Journal of Molecular Sciences·Ariel KantorRobert E MacLaren
Feb 28, 2017·BMC Medicine·Jessica L SchnellerCharles P Venditti
Dec 7, 2018·Genome Biology·Nadia AmraniErik J Sontheimer
Mar 19, 2019·Annual Review of Biochemistry·Daesik KimJin-Soo Kim
Jul 15, 2020·Nature Biotechnology·Volker WiebkingMatthew H Porteus
Apr 5, 2016·Oral Diseases·A Peterson
Jun 22, 2017·Mammalian Genome : Official Journal of the International Mammalian Genome Society·Alberto Cebrian-Serrano, Benjamin Davies

❮ Previous
Next ❯

Methods Mentioned

BETA
transfection
transfections
PCR

Software Mentioned

COSMID
binom package
R
Nme

Related Concepts

Related Feeds

CRISPR (general)

Clustered regularly interspaced short palindromic repeats (CRISPR) are DNA sequences in the genome that are recognized and cleaved by CRISPR-associated proteins (Cas). CRISPR-Cas system enables the editing of genes to create or correct mutations. Discover the latest research on CRISPR here.

CRISPR for Genome Editing

Genome editing technologies enable the editing of genes to create or correct mutations. Clustered regularly interspaced short palindromic repeats (CRISPR) are DNA sequences in the genome that are recognized and cleaved by CRISPR-associated proteins (Cas). Here is the latest research on the use of CRISPR-Cas system in gene editing.

CRISPR Ribonucleases Deactivation

CRISPR-Cas system enables the editing of genes to create or correct mutations. This feed focuses on mechanisms that underlie deactivation of CRISPR ribonucleases. Here is the latest research.