A synthetic oscillatory network of transcriptional regulators

Networks of interacting biomolecules carry out many essential functions in living cells(1), but the 'design principles' underlying the functioning of such intracellular networks remain poorly understood, despite intensive eff orts including quantitative analysis of relatively simple systems(2). Here we present a complementary approach to this problem: the design and constru ction of a synthetic network to implement a particular function. We used th ree transcriptional repressor systems that are not part of any natural biol ogical clock(3-5) to build an oscillating network, termed the repressilator , in Escherichia coli. The network periodically induces the synthesis of gr een fluorescent protein as a readout of its state in individual cells. The resulting oscillations, with typical periods of hours, are slower than the cell-division cycle, so the state of the oscillator has to be transmitted f rom generation to generation. This artificial clock displays noisy behaviou r, possibly because of stochastic fluctuations of its components. Such 'rat ional network design' may lead both to the engineering of new cellular beha viours and to an improved understanding of naturally occurring networks.