title: Applications of Cas9 as an RNA-programmed RNA-binding protein.
abstract: The Streptococcus pyogenes CRISPR-Cas system has gained widespread application as a genome editing and gene regulation tool as simultaneous cellular delivery of the Cas9 protein and guide RNAs enables recognition of specific DNA sequences. The recent discovery that Cas9 can also bind and cleave RNA in an RNA-programmable manner indicates the potential utility of this system as a universal nucleic acid-recognition technology. RNA-targeted Cas9 (RCas9) could allow identification and manipulation of RNA substrates in live cells, empowering the study of cellular gene expression, and could ultimately spawn patient- and disease-specific diagnostic and therapeutic tools. Here we describe the development of RCas9 and compare it to previous methods for RNA targeting, including engineered RNA-binding proteins and other types of CRISPR-Cas systems. We discuss potential uses ranging from live imaging of transcriptional dynamics to patient-specific therapies and applications in synthetic biology.
authors.affiliation: Department of Cellular and Molecular Medicine, Stem Cell Program, and Institute for Genomic Medicine, University of California San Diego, La Jolla, CA, USA.
authors.forename: David A
authors.initials: DA
authors.lastname: Nelles
authors.affiliation: Department of Cellular and Molecular Medicine, Stem Cell Program, and Institute for Genomic Medicine, University of California San Diego, La Jolla, CA, USA.
authors.forename: Mark Y
authors.initials: MY
authors.lastname: Fang
authors.affiliation: Department of Cellular and Molecular Medicine, Stem Cell Program, and Institute for Genomic Medicine, University of California San Diego, La Jolla, CA, USA.
authors.forename: Stefan
authors.initials: S
authors.lastname: Aigner
authors.affiliation: Department of Cellular and Molecular Medicine, Stem Cell Program, and Institute for Genomic Medicine, University of California San Diego, La Jolla, CA, USA.|Molecular Engineering Laboratory, Biomedical Sciences Institutes, Agency for Science, Technology & Research and Yong Loo Lin School of Medicine, National University of Singapore, Singapore.
authors.forename: Gene W
authors.initials: GW
authors.lastname: Yeo
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics*; D000076987:CRISPR-Associated Protein 9; D064113:CRISPR-Cas Systems; D004720:Endonucleases / Q000235:genetics*; D005786:Gene Expression Regulation; D005818:Genetic Engineering; D006801:Humans; D016601:RNA-Binding Proteins / Q000235:genetics*; D058615:Synthetic Biology
chemical_list: D001426:Bacterial Proteins; D016601:RNA-Binding Proteins; D000076987:CRISPR-Associated Protein 9; C000600049:Cas9 protein, Francisella novicida; D004720:Endonucleases
keywords: CRISPR-Cas; Cas9; RCas9; RNA biology; RNA targeting; RNA-binding proteins