A Choline-Responsive Band-Pass gene circuit without transcriptional cascades.

Band-pass filters, which selectively transmit signals within a defined range of input magnitudes, are fundamental components of signal-processing systems. In cellular gene circuits, band-pass behavior has likewise been pursued as a mean to implement complex signal-processing functions. However, previously reported genetic band-pass circuits have typically relied not only on a large number of regulatory components but also on transcriptional cascades involving multiple transcription factors, resulting in long DNA sequences and increased circuit complexity. Here, we first propose a band-pass gene circuit that operates without transcriptional cascades. By co-expressing two variants of the transcription factor BetI that exhibit opposite input-response behaviors-one acting as an inducer-dependent activator and the other as an inducer-dependent repressor-band-pass filtering is achieved solely through differential tuning of their inducer sensitivities. This minimal architecture enables gene expression only within a specific range of intracellular choline concentrations. Furthermore, we demonstrate that this cascade-free band-pass circuit can be exploited to generate spatial expression patterns in Escherichia coli populations in response to a choline diffusion gradient, illustrating its utility for pattern formation in multicellular contexts.