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ACS Synth Biol. 2014 Dec 19;3(12):929-31. doi: 10.1021/sb500036q. Epub 2014 Dec 6.

Specific gene repression by CRISPRi system transferred through bacterial conjugation.

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Center for Systems and Synthetic Biology, ‡Department of Cellular and Molecular Pharmacology, §Department of Biochemistry and Biophysics, ∥Department of Bioengineering and Therapeutic Sciences, ⊥Howard Hughes Medical Institute, University of California, San Francisco , San Francisco, California 94158, United States.


In microbial communities, bacterial populations are commonly controlled using indiscriminate, broad range antibiotics. There are few ways to target specific strains effectively without disrupting the entire microbiome and local environment. Here, we use conjugation, a natural DNA horizontal transfer process among bacterial species, to deliver an engineered CRISPR interference (CRISPRi) system for targeting specific genes in recipient Escherichia coli cells. We show that delivery of the CRISPRi system is successful and can specifically repress a reporter gene in recipient cells, thereby establishing a new tool for gene regulation across bacterial cells and potentially for bacterial population control.


CRISPR/Cas9; conjugation; horizontal gene transfer; synthetic biology; synthetic gene regulation

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