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Nucleic acids research DOI:10.1093/nar/gkt520

Programmable repression and activation of bacterial gene expression using an engineered CRISPR-Cas system.

Publication TypeJournal Article
Year of Publication2013
AuthorsBikard, D, Jiang, W, Samai, P, Hochschild, A, Zhang, F, Marraffini, LA
JournalNucleic acids research
Date Published2013/06/12

The ability to artificially control transcription is essential both to the study of gene function and to the construction of synthetic gene networks with desired properties. Cas9 is an RNA-guided double-stranded DNA nuclease that participates in the CRISPR-Cas immune defense against prokaryotic viruses. We describe the use of a Cas9 nuclease mutant that retains DNA-binding activity and can be engineered as a programmable transcription repressor by preventing the binding of the RNA polymerase (RNAP) to promoter sequences or as a transcription terminator by blocking the running RNAP. In addition, a fusion between the omega subunit of the RNAP and a Cas9 nuclease mutant directed to bind upstream promoter regions can achieve programmable transcription activation. The simple and efficient modulation of gene expression achieved by this technology is a useful asset for the study of gene networks and for the development of synthetic biology and biotechnological applications.