Orthogonal Cas9 proteins for RNA-guided gene regulation and editing
A set of Cas9 endonucleases orthogonal to the Streptococcus pyogenes enzyme is identified. This will enable simultaneous addressing of multiple RNA-guided activities to different genomic target sites with the CRISPR-Cas9 system. The Cas9 protein from the Streptococcus pyogenes CRISPR-Cas acquired im...
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| Veröffentlicht in: | Nature methods 2013-11, Vol.10 (11), p.1116-1121 |
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| creator | Esvelt, Kevin M Mali, Prashant Braff, Jonathan L Moosburner, Mark Yaung, Stephanie J Church, George M |
| description | A set of Cas9 endonucleases orthogonal to the
Streptococcus pyogenes
enzyme is identified. This will enable simultaneous addressing of multiple RNA-guided activities to different genomic target sites with the CRISPR-Cas9 system.
The Cas9 protein from the
Streptococcus pyogenes
CRISPR-Cas acquired immune system has been adapted for both RNA-guided genome editing and gene regulation in a variety of organisms, but it can mediate only a single activity at a time within any given cell. Here we characterize a set of fully orthogonal Cas9 proteins and demonstrate their ability to mediate simultaneous and independently targeted gene regulation and editing in bacteria and in human cells. We find that Cas9 orthologs display consistent patterns in their recognition of target sequences, and we identify an unexpectedly versatile Cas9 protein from
Neisseria meningitidis
. We provide a basal set of orthogonal RNA-guided proteins for controlling biological systems and establish a general methodology for characterizing additional proteins. |
| doi_str_mv | 10.1038/nmeth.2681 |
| format | Article |
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Streptococcus pyogenes
enzyme is identified. This will enable simultaneous addressing of multiple RNA-guided activities to different genomic target sites with the CRISPR-Cas9 system.
The Cas9 protein from the
Streptococcus pyogenes
CRISPR-Cas acquired immune system has been adapted for both RNA-guided genome editing and gene regulation in a variety of organisms, but it can mediate only a single activity at a time within any given cell. Here we characterize a set of fully orthogonal Cas9 proteins and demonstrate their ability to mediate simultaneous and independently targeted gene regulation and editing in bacteria and in human cells. We find that Cas9 orthologs display consistent patterns in their recognition of target sequences, and we identify an unexpectedly versatile Cas9 protein from
Neisseria meningitidis
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Streptococcus pyogenes
enzyme is identified. This will enable simultaneous addressing of multiple RNA-guided activities to different genomic target sites with the CRISPR-Cas9 system.
The Cas9 protein from the
Streptococcus pyogenes
CRISPR-Cas acquired immune system has been adapted for both RNA-guided genome editing and gene regulation in a variety of organisms, but it can mediate only a single activity at a time within any given cell. Here we characterize a set of fully orthogonal Cas9 proteins and demonstrate their ability to mediate simultaneous and independently targeted gene regulation and editing in bacteria and in human cells. We find that Cas9 orthologs display consistent patterns in their recognition of target sequences, and we identify an unexpectedly versatile Cas9 protein from
Neisseria meningitidis
. 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Academic</collection><collection>Nucleic Acids Abstracts</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Nature methods</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Esvelt, Kevin M</au><au>Mali, Prashant</au><au>Braff, Jonathan L</au><au>Moosburner, Mark</au><au>Yaung, Stephanie J</au><au>Church, George M</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Orthogonal Cas9 proteins for RNA-guided gene regulation and editing</atitle><jtitle>Nature methods</jtitle><stitle>Nat Methods</stitle><addtitle>Nat Methods</addtitle><date>2013-11-01</date><risdate>2013</risdate><volume>10</volume><issue>11</issue><spage>1116</spage><epage>1121</epage><pages>1116-1121</pages><issn>1548-7091</issn><eissn>1548-7105</eissn><abstract>A set of Cas9 endonucleases orthogonal to the
Streptococcus pyogenes
enzyme is identified. This will enable simultaneous addressing of multiple RNA-guided activities to different genomic target sites with the CRISPR-Cas9 system.
The Cas9 protein from the
Streptococcus pyogenes
CRISPR-Cas acquired immune system has been adapted for both RNA-guided genome editing and gene regulation in a variety of organisms, but it can mediate only a single activity at a time within any given cell. Here we characterize a set of fully orthogonal Cas9 proteins and demonstrate their ability to mediate simultaneous and independently targeted gene regulation and editing in bacteria and in human cells. We find that Cas9 orthologs display consistent patterns in their recognition of target sequences, and we identify an unexpectedly versatile Cas9 protein from
Neisseria meningitidis
. We provide a basal set of orthogonal RNA-guided proteins for controlling biological systems and establish a general methodology for characterizing additional proteins.</abstract><cop>New York</cop><pub>Nature Publishing Group US</pub><pmid>24076762</pmid><doi>10.1038/nmeth.2681</doi><tpages>6</tpages><orcidid>https://orcid.org/0000-0001-8797-3945</orcidid><orcidid>https://orcid.org/0000000187973945</orcidid><oa>free_for_read</oa></addata></record> |
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| subjects | 631/1647/1511 631/1647/338/552 631/208/200 631/337/572 Amino Acid Sequence Analysis Bacterial Proteins - chemistry Bacterial Proteins - genetics Bacterial Proteins - physiology Bioinformatics Biological Microscopy Biological Techniques Biomedical Engineering/Biotechnology Cells Gene Expression Regulation Gene mutations Genes Genetic regulation Immune system Life Sciences Molecular Sequence Data Neisseria meningitidis Protein research Proteins Proteomics Ribonucleic acid RNA RNA - genetics RNA Editing Sequence Homology, Amino Acid Streptococcus pyogenes Streptomyces - physiology |
| title | Orthogonal Cas9 proteins for RNA-guided gene regulation and editing |
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