Large-scale design of robust genetic circuits with multiple inputs and outputs for mammalian cells

Genetic circuits that are reliable, robust, and scalable are built without the need for optimization using a recombinase-based system. Engineered genetic circuits for mammalian cells often require extensive fine-tuning to perform as intended. We present a robust, general, scalable system, called �...

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Bibliographic Details
Published in:Nature biotechnology 2017-05, Vol.35 (5), p.453-462
Main Authors: Weinberg, Benjamin H, Pham, N T Hang, Caraballo, Leidy D, Lozanoski, Thomas, Engel, Adrien, Bhatia, Swapnil, Wong, Wilson W
Format: Article
Language:English
Subjects:
38/35
631/553/2691
631/553/552
96/106
Agriculture
Bioinformatics
Biomedical Engineering/Biotechnology
Biomedicine
Biotechnology
Cellular Reprogramming Techniques - methods
Computer Simulation
Computers, Molecular
Design
Eukaryotes
Gene Regulatory Networks - genetics
Genetic engineering
Genetic Engineering - methods
Genetic research
Humans
Jurkat Cells
Life Sciences
Mammals
Methods
Models, Genetic
Prokaryotes
Proteome - genetics
Signal Transduction - genetics
Synthetic biology
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Summary:Genetic circuits that are reliable, robust, and scalable are built without the need for optimization using a recombinase-based system. Engineered genetic circuits for mammalian cells often require extensive fine-tuning to perform as intended. We present a robust, general, scalable system, called 'Boolean logic and arithmetic through DNA excision' (BLADE), to engineer genetic circuits with multiple inputs and outputs in mammalian cells with minimal optimization. The reliability of BLADE arises from its reliance on recombinases under the control of a single promoter, which integrates circuit signals on a single transcriptional layer. We used BLADE to build 113 circuits in human embryonic kidney and Jurkat T cells and devised a quantitative, vector-proximity metric to evaluate their performance. Of 113 circuits analyzed, 109 functioned (96.5%) as intended without optimization. The circuits, which are available through Addgene, include a 3-input, two-output full adder; a 6-input, one-output Boolean logic look-up table; circuits with small-molecule-inducible control; and circuits that incorporate CRISPR–Cas9 to regulate endogenous genes. BLADE enables execution of sophisticated cellular computation in mammalian cells, with applications in cell and tissue engineering.
ISSN:1087-0156
1546-1696
DOI:10.1038/nbt.3805