title: Orthogonal gene knockout and activation with a catalytically active Cas9 nuclease.
abstract: We have developed a CRISPR-based method that uses catalytically active Cas9 and distinct single guide (sgRNA) constructs to knock out and activate different genes in the same cell. These sgRNAs, with 14- to 15-bp target sequences and MS2 binding loops, can activate gene expression using an active Streptococcus pyogenes Cas9 nuclease, without inducing double-stranded breaks. We use these 'dead RNAs' to perform orthogonal gene knockout and transcriptional activation in human cells.
authors.forename: James E
authors.initials: JE
authors.lastname: Dahlman
authors.forename: Omar O
authors.initials: OO
authors.lastname: Abudayyeh
authors.forename: Julia
authors.initials: J
authors.lastname: Joung
authors.forename: Jonathan S
authors.initials: JS
authors.lastname: Gootenberg
authors.forename: Feng
authors.initials: F
authors.lastname: Zhang
authors.forename: Silvana
authors.initials: S
authors.lastname: Konermann
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics*; D000076987:CRISPR-Associated Protein 9; D004720:Endonucleases / Q000235:genetics*; D055786:Gene Knockout Techniques / Q000379:methods*; D057809:HEK293 Cells; D006801:Humans; D000094704:RNA, Guide, CRISPR-Cas Systems / Q000235:genetics*
chemical_list: D001426:Bacterial Proteins; D000094704:RNA, Guide, CRISPR-Cas Systems; D000076987:CRISPR-Associated Protein 9; C000606107:Cas9 endonuclease Streptococcus pyogenes; D004720:Endonucleases