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. 2017 May 25;8:15459. doi: 10.1038/ncomms15459

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Copyright © 2017, The Author(s)

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(a) A NOR gate input stage consists of a Pol II pGRR promoter that is fully repressed by the binding of either one or both of its cognate gRNA-dCas9-Mxi1 complexes. The output stage of the NOR gate is a gRNA transcript, flanked by self-cleaving ribozymes (RGR). Cleavage sites are indicated by red arrows. The cleavage of the ribozymes prevents nuclear export of the gRNA, indicated by dotted grey arrow. (b) The process of NOR gate library construction. Our library consists of a set of 400 two-input pGRR promoters and 20 RGR outputs for a total of 8,000 possible NOR gates. (c) Genomically integrating NOR gates into S. cerevisiae. (d) Arbitrary circuits are constructed by integrating multiple NOR gates into a single strain.