Details:

Alternative Title:
Nucleic Acids Res
Publisher's site:
http://dx.doi.org/10.1093/nar/gkt520
Personal Author:
Bikard, David ; Jiang, Wenyan ; Samai, Poulami ; Hochschild, Ann ; Zhang, Feng ; ... More +
Bikard, David ; Jiang, Wenyan ; Samai, Poulami ; Hochschild, Ann ; Zhang, Feng ; Marraffini, Luciano A. Less -
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.
Subjects:
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Bacterial Proteins Endoribonucleases Escherichia Coli Gene Expression Regulation, Bacterial Gene Regulatory Networks Genes, Bacterial Genes, Synthetic Genetic Loci Inverted Repeat Sequences Molecular Biology Promoter Regions, Genetic Protein Binding Repressor Proteins RNA, Bacterial Streptococcus Pneumoniae Streptococcus Pyogenes Transcriptional Activation
Source:
Nucleic Acids Res. 2013; 41(15):7429-7437.
Pubmed ID:
23761437
Pubmed Central ID:
PMC3753641
Document Type:
Journal Article
Funding:
1DP2AI104556-01/AI/NIAID NIH HHS/United States ; DP1MH100706/DP/NCCDPHP CDC HHS/United States ; DP2 AI104556/AI/NIAID NIH HHS/United States ; R01 GM044025/GM/NIGMS NIH HHS/United States ; R01 GM044025/GM/NIGMS NIH HHS/United States
1DP2AI104556-01/AI/NIAID NIH HHS/United States ; DP1MH100706/DP/NCCDPHP CDC HHS/United States ; DP2 AI104556/AI/NIAID NIH HHS/United States ; R01 GM044025/GM/NIGMS NIH HHS/United States ; R01 GM044025/GM/NIGMS NIH HHS/United States Less -
Collection(s):
CDC Public Access
Main Document Checksum:
[+]
urn:sha256:9bfe3d623c18094620d12fb2e18e2a772f6fb703bd355273efa15c30cc5d3d83
Download URL:
https://stacks.cdc.gov/view/cdc/22580/cdc_22580_DS1.pdf
File Type:

Supporting Files

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