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. 2008 Oct 28;4:224. doi: 10.1038/msb.2008.62

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Copyright © 2008, EMBO and Nature Publishing Group

This is an open-access article distributed under the terms of the Creative Commons Attribution Licence, which permits distribution and reproduction in any medium, provided the original author and source are credited. Creation of derivative works is permitted but the resulting work may be distributed only under the same or similar licence to this one. This licence does not permit commercial exploitation without specific permission.

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Programming transfer functions through combinatorial design strategies. (A) Combinatorial tuning strategies enable fine-tuning of the component transfer function. Stepwise nucleotide changes were made to S4, where each change fell under a different competing strand tuning strategy. (B) Tuned theophylline response curves as described in Figure 1D. Arrows depict the systematic modifications designated in (A). (C) Circuit configuration of shRNA switches responsive to theophylline (S4) or hypoxanthine (X1) that both target EGFP. (D) Predicted transfer function on the basis of application of the mathematical model to the circuit depicted in (C). Fit curves represent the individual component transfer functions for S4 () and X1 (), respectively. (E) Combinatorial implementation of shRNA switches enables construction of networks that process multiple molecular inputs. Results are shown for HEK293T tTA-d2EGFP cells transfected with each shRNA construct (250 ng) or cotransfected with both shRNA constructs (125 ng of each) in the presence of water (▪), 3 mM theophylline (), 2 mM hypoxanthine (), or both theophylline and hypoxanthine (□). Error bars represent one s.d. from duplicate transfected wells.

Inline graphic — Programming transfer functions through combinatorial design strategies. (A) Combinatorial tuning strategies enable fine-tuning of the component transfer function. Stepwise nucleotide changes were made to S4, where each change fell under a different competing strand tuning strategy. (B) Tuned theophylline response curves as described in Figure 1D. Arrows depict the systematic modifications designated in (A). (C) Circuit configuration of shRNA switches responsive to theophylline (S4) or hypoxanthine (X1) that both target EGFP. (D) Predicted transfer function on the basis of application of the mathematical model to the circuit depicted in (C). Fit curves represent the individual component transfer functions for S4 () and X1 (), respectively. (E) Combinatorial implementation of shRNA switches enables construction of networks that process multiple molecular inputs. Results are shown for HEK293T tTA-d2EGFP cells transfected with each shRNA construct (250 ng) or cotransfected with both shRNA constructs (125 ng of each) in the presence of water (▪), 3 mM theophylline (), 2 mM hypoxanthine (), or both theophylline and hypoxanthine (□). Error bars represent one s.d. from duplicate transfected wells.