ORIGINAL: NATURE BIOTECHNOLOGY | RESEARCH | LETTER
Corresponding author Nature Biotechnology (2013) doi:10.1038/nbt.2510Received 25 October 2012 Accepted 17 January 2013 Published online 10 February 2013
Logic and memory are essential functions of circuits that generate complex, state-dependent responses. Here we describe a strategy for efficiently assembling synthetic genetic circuits that use recombinases to implement Boolean logic functions with stable DNA-encoded memory of events.
Application of this strategy allowed us to create all 16 two-input Boolean logic functions in living Escherichia coli cells without requiring cascades comprising multiple logic gates. We demonstrate long-term maintenance of memory for at least 90 cell generations and the ability to interrogate the states of these synthetic devices with fluorescent reporters and PCR.
Using this approach we created two-bit digital-to-analog converters, which should be useful in biotechnology applications for encoding multiple stable gene expression outputs using transient inputs of inducers. We envision that this integrated logic and memory system will enable the implementation of complex cellular state machines, behaviors and pathways for therapeutic, diagnostic and basic science applications.
At a glance
Figure 2: Recombinase-based logic gates can implement a complete set of two-input–one-output Boolean logic gates without needing to cascade multiple universal gates together. |
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Department of Electrical Engineering and Computer Science, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
Piro Siuti &
Timothy K Lu
Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
Piro Siuti &
Timothy K Lu
Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
John Yazbek
Contributions
T.K.L. conceived of this study. P.S. and J.Y. implemented, constructed and performed all experiments. All authors analyzed the data, discussed results and wrote the manuscript.
Competing financial interests
P.S., J.Y. and T.K.L. have filed a provisional application with the US Patent and Trademark Office on this work.
Corresponding author
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Supplementary information
PDF files
Supplementary Figures 1–4, Supplementary Table 1 and Supplementary Data
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