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Nucleic Acids Res DOI:10.1093/nar/gkt520

Programmable repression and activation of bacterial gene expression using an engineered CRISPR-Cas system.

Publication TypeJournal Article
Year of Publication2013
AuthorsBikard, D, Jiang, W, Samai, P, Hochschild, A, Zhang, F, Marraffini, LA
JournalNucleic Acids Res
Date Published2013 Aug
KeywordsBacterial Proteins, Endoribonucleases, Escherichia coli, Gene Expression Regulation, Bacterial, Gene Regulatory Networks, Genes, Bacterial, Genes, Synthetic, Genetic Loci, Inverted Repeat Sequences, Promoter Regions, Genetic, Protein Binding, Repressor Proteins, RNA, Bacterial, Streptococcus pneumoniae, Streptococcus pyogenes, Transcriptional Activation

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.


Alternate JournalNucleic Acids Res.
PubMed ID23761437
PubMed Central IDPMC3753641
Grant ListDP2 AI104556 / AI / NIAID 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