Details

Autor(en) / Beteiligte

• Bikard, David
• Jiang, Wenyan
• Samai, Poulami
• Hochschild, Ann
• Zhang, Feng
• Marraffini, Luciano A

Titel

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

Ist Teil von

• Nucleic acids research, 2013-08, Vol.41 (15), p.7429-7437

Ort / Verlag

England

Erscheinungsjahr

2013

Quelle

MEDLINE

Beschreibungen/Notizen

• 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.