Programmable repression and activation of bacterial gene expression using an engineered CRISPR-Cas system
Name
Bikard-2013-Programmable repress.pdf
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3.05 MB
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Adobe PDF
Checksum (MD5)
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Author(s)
Zhang, Feng
Bikard, David
Jiang, Wenyan
Samai, Poulami
Hochschild, Ann
Marraffini, Luciano A.
Date Issued
June 2013
Journal
Nucleic Acids Research
Publisher
Oxford University Press
Citation
Bikard, D. et al. “Programmable Repression and Activation of Bacterial Gene Expression Using an Engineered CRISPR-Cas System.” Nucleic Acids Research (2013).
Version
Final published version
Abstract
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.
MIT Department
Massachusetts Institute of Technology. Department of Biological Engineering
Massachusetts Institute of Technology. Department of Brain and Cognitive Sciences
McGovern Institute for Brain Research at MIT
Terms of Use
Creative Commons Attribution Non-Commercial
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DOI of Published Version
https://doi.org/10.1093/nar/gkt520
