Programmable repression and activation of bacterial gene expression using an engineered CRISPR-Cas system
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 viru...
| Main Authors: | , , , , , |
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| Other Authors: | |
| Format: | Article |
| Language: | en_US |
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Oxford University Press
2013
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| Online Access: | http://hdl.handle.net/1721.1/79745 https://orcid.org/0000-0003-2782-2509 |
| _version_ | 1826189457872650240 |
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| author | Zhang, Feng Bikard, David Jiang, Wenyan Samai, Poulami Hochschild, Ann Marraffini, Luciano A. |
| author2 | Massachusetts Institute of Technology. Department of Biological Engineering |
| author_facet | Massachusetts Institute of Technology. Department of Biological Engineering Zhang, Feng Bikard, David Jiang, Wenyan Samai, Poulami Hochschild, Ann Marraffini, Luciano A. |
| author_sort | Zhang, Feng |
| collection | MIT |
| description | 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. |
| first_indexed | 2024-09-23T08:15:22Z |
| format | Article |
| id | mit-1721.1/79745 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T08:15:22Z |
| publishDate | 2013 |
| publisher | Oxford University Press |
| record_format | dspace |
| spelling | mit-1721.1/797452022-09-30T08:37:33Z Programmable repression and activation of bacterial gene expression using an engineered CRISPR-Cas system Zhang, Feng Bikard, David Jiang, Wenyan Samai, Poulami Hochschild, Ann Marraffini, Luciano A. 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 Zhang, Feng 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. National Institutes of Health (U.S.) (Pioneer Award DP1MH100706) National Institutes of Health (U.S.) (Transformative Research Award) W. M. Keck Foundation McKnight Foundation Bill & Melinda Gates Foundation Damon Runyon Cancer Research Foundation Kinship Foundation. Searle Scholars Program Esther A. & Joseph Klingenstein Fund, Inc. Simons Foundation 2013-07-31T17:56:54Z 2013-07-31T17:56:54Z 2013-06 2013-05 Article http://purl.org/eprint/type/JournalArticle 0305-1048 1362-4962 http://hdl.handle.net/1721.1/79745 Bikard, D. et al. “Programmable Repression and Activation of Bacterial Gene Expression Using an Engineered CRISPR-Cas System.” Nucleic Acids Research (2013). https://orcid.org/0000-0003-2782-2509 en_US http://dx.doi.org/10.1093/nar/gkt520 Nucleic Acids Research Creative Commons Attribution Non-Commercial http://creativecommons.org/licenses/by-nc/3.0 application/pdf Oxford University Press Oxford University Press |
| spellingShingle | Zhang, Feng Bikard, David Jiang, Wenyan Samai, Poulami Hochschild, Ann Marraffini, Luciano A. Programmable repression and activation of bacterial gene expression using an engineered CRISPR-Cas system |
| title | Programmable repression and activation of bacterial gene expression using an engineered CRISPR-Cas system |
| title_full | Programmable repression and activation of bacterial gene expression using an engineered CRISPR-Cas system |
| title_fullStr | Programmable repression and activation of bacterial gene expression using an engineered CRISPR-Cas system |
| title_full_unstemmed | Programmable repression and activation of bacterial gene expression using an engineered CRISPR-Cas system |
| title_short | Programmable repression and activation of bacterial gene expression using an engineered CRISPR-Cas system |
| title_sort | programmable repression and activation of bacterial gene expression using an engineered crispr cas system |
| url | http://hdl.handle.net/1721.1/79745 https://orcid.org/0000-0003-2782-2509 |
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