Programmable repression and activation of bacterial gene expression using an engineered CRISPR-Cas system
Author(s)Zhang, Feng; Bikard, David; Jiang, Wenyan; Samai, Poulami; Hochschild, Ann; Marraffini, Luciano A.; ... Show more Show less
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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.
DepartmentMassachusetts Institute of Technology. Department of Biological Engineering; Massachusetts Institute of Technology. Department of Brain and Cognitive Sciences; McGovern Institute for Brain Research at MIT
Nucleic Acids Research
Oxford University Press
Bikard, D. et al. “Programmable Repression and Activation of Bacterial Gene Expression Using an Engineered CRISPR-Cas System.” Nucleic Acids Research (2013).
Final published version