Fig. 3. PERSIST endoRNase platform is modular.

All 16 two-input Boolean logic functions can be created using Cas endoRNases and the PERSIST-ON- and OFF-switches. Each logic function was constructed using EYFP as output with other constructs included as necessary to confer a designed response to two inputs: Csy4 and CasE. Cse3 endoRNase was used as an intermediate regulator where necessary. Each set of constructs encoding specific logic functions was evaluated without input endoRNases, with CasE only, with Csy4 only, and with both Csy4 and CasE via poly-transfection (Supplementary Fig. 15). The expected Boolean logic output is depicted below each chart. All logic functions can be correctly identified using the same approximately 10-fold separation between ON and OFF values (gray region). n = 2 or n = 3 biologically independent samples as indicated by the points where each sample represents the evaluation of >1000 transfected cells (HEK293FT). Data bars are presented as mean normalized output.