Multi-input CRISPR/Cas genetic circuits that interface host regulatory networks

摘要: Genetic circuits require many regulatory parts in order to implement signal processing or execute algorithms cells. A potentially scalable approach is use dCas9, which employs small guide RNAs (sgRNAs) repress genetic loci via the programmability of RNA:DNA base pairing. To this end, we dCas9 and designed sgRNAs build transcriptional logic gates connect them perform computation living We constructed a set NOT by designing five synthetic Escherichia coli σ70 promoters that are repressed corresponding sgRNAs, these interactions do not exhibit crosstalk between each other. These high on-target repression (56- 440-fold) negligible off-target (< 1.3-fold). were connected larger circuits, including Boolean-complete NOR gate 3-gate circuit consisting four layered sgRNAs. The native E. network output target an transcription factor (malT). This converts switch cellular phenotype (sugar utilization, chemotaxis, phage resistance).