Rationally designed logic integration of regulatory signals in mammalian cells.
Molecular-level information processing is essential for 'smart' in vivo nanosystems. Natural molecular computing, such as the regulation of messenger RNA (mRNA) synthesis by special proteins called transcription factors, has inspired engineered systems 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 output. We construct, test and optimize increasingly complex circuits with up to three transcription factor inputs, establishing a platform for in vivo molecular computing.
Leisner
Madeleine
M
FAS Centre for Systems Biology, Harvard University, 52 Oxford Street, Cambridge Massachussetts 02138 USA.
Bleris
Leonidas
L
Lohmueller
Jason
J
Xie
Zhen
Z
Benenson
Yaakov
Y
D000818:Animals; D002460:Cell Line; D039301:Computers, Molecular; D005786:Gene Expression Regulation; D006801:Humans; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism; D008957:Models, Genetic; D010957:Plasmids / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*; D014162:Transfection
D035683:MicroRNAs; D034741:RNA, Small Interfering; D014157:Transcription Factors