Journal
NUCLEIC ACIDS RESEARCH
Volume 41, Issue 15, Pages 7429-7437Publisher
OXFORD UNIV PRESS
DOI: 10.1093/nar/gkt520
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Funding
- Harvey L. Karp Discovery Award
- Bettencourt Schuller Foundation
- Helmsley Postdoctoral Fellowship for Basic and Translational Research on Disorders of the Digestive System at The Rockefeller University
- NIH [R01 GM044025]
- NIH Director's Pioneer Award [DP1MH100706]
- Transformative R01 Foundation
- Keck Foundation
- McKnight Foundation
- Gates Foundation
- Damon Runyon Foundation
- Searle Scholars Foundation
- Klingenstein Foundation
- Simons Foundation
- Searle Scholars Program
- Rita Allen Scholars Program
- Irma T. Hirschl Award
- NIH Director's New Innovator Award [1DP2AI104556-01]
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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.
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