Biological 2-input decoder circuit in human cells.
Decoders are combinational circuits that convert information from n inputs to a maximum of 2(n) outputs. This operation is of major importance in computing systems yet it is vastly underexplored in synthetic biology. Here, we present a synthetic gene network architecture that operates as a biological decoder in human cells, converting 2 inputs to 4 outputs. As a proof-of-principle, we use small molecules to emulate the two inputs and fluorescent reporters as the corresponding four outputs. The experiments are performed using transient transfections in human kidney embryonic cells and the characterization by fluorescence microscopy and flow cytometry. We show a clear separation between the ON and OFF mean fluorescent intensity states. Additionally, we adopt the integrated mean fluorescence intensity for the characterization of the circuit and show that this metric is more robust to transfection conditions when compared to the mean fluorescent intensity. To conclude, we present the first implementation of a genetic decoder. This combinational system can be valuable toward engineering higher-order circuits as well as accommodate a multiplexed interface with endogenous cellular functions.
Bioengineering Department, The University of Texas at Dallas , 800 West Campbell Road, Richardson, Texas 75080, United States.
Michael
M
Guinn
Leonidas
L
Bleris
D002478:Cells, Cultured; D005456:Fluorescent Dyes / Q000032:analysis* / Q000378:metabolism; D053263:Gene Regulatory Networks; D005813:Genes, Synthetic; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007668:Kidney / Q000166:cytology / Q000196:embryology; D008962:Models, Theoretical; D011401:Promoter Regions, Genetic; D058615:Synthetic Biology / Q000379:methods*
D005456:Fluorescent Dyes; D049452:Green Fluorescent Proteins