Abstract
Molecular-level information processing1,2 is essential for ‘smart’ in vivo nanosystems. Natural molecular computing, such as the regulation of messenger RNA (mRNA) synthesis by special proteins called transcription factors3,4, has inspired engineered systems5,6,7,8,9,10,11,12,13,14,15 that can control the levels of mRNA with certain combinations of transcription factors. Here, we show an alternative approach to achieving general-purpose control of mRNA and protein levels by logic integration of transcription factor input signals in mammalian cells. The transcription factors regulate synthetic genes coding for small regulatory RNAs (called microRNAs), which, in turn, control the mRNA of interest (the output) via an RNA interference pathway. The simplicity of these modular interactions makes it possible, in theory, to implement any arbitrary logic relation between the transcription factors and the output16. We construct, test and optimize increasingly complex circuits with up to three transcription factor inputs, establishing a platform for in vivo molecular computing.
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Acknowledgements
The authors would like to thank anonymous reviewers for thoughtful comments. M.L. is a recipient of Deutsche Forschungs Gemeinschaft scholarship. The research was funded by the Bauer Fellows program and by the NIGMS grant GM068763 for National Centres of Systems Biology. M.L. acknowledges her father R. Leisner, who passed away as this manuscript was prepared for submission: ‘One learns most from those one loves’ (Goethe).
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Y.B. designed the research and supervised the project. M.L., L.B., J.L., Z.X. and Y.B. performed the research. M.L., Y.B., J.L. and L.B. wrote the manuscript.
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Leisner, M., Bleris, L., Lohmueller, J. et al. Rationally designed logic integration of regulatory signals in mammalian cells. Nature Nanotech 5, 666–670 (2010). https://doi.org/10.1038/nnano.2010.135
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DOI: https://doi.org/10.1038/nnano.2010.135
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