Antisense RNA interactions are present in all domains of life. Typically sense-antisense RNA pairs originate from overlapping genes with convergent face to face promoters, and are speculated to be involved in gene regulation and development of life-threatening diseases in humans Recent studies indicate the role of transcriptional interference in regulating expression of genes in convergent orientation. We present a mathematical modeling and experimental framework to understand convergent transcription that combines the effects of transcriptional interference (TI) and cis-antisense RNA based regulation (AR). Our model shows that combination of TI and AR adds multiple-levels of regulation which allows such a system to have a highly tunable output in response to stimuli, ranging from a simple-first order response to biologically complex higher-order response such as bistable switch. We demonstrate a library of experimental constructs with engineered TI and AR by using face-to-face inducible promoters separated by carefully tailored overlapping DNA sequences to control expression of a set of fluorescent reporter proteins. We show that engineered TI and AR allows for design of a highly-tunable synthetic genetic switch. Studying these coupled gene expression mechanisms allows for an understanding of higher-order behavior of regulatory gene expression networks and design of novel synthetic genetic switch devices. Combining multiple switch devices that function differently depending on environmental conditions allows for creation of complex biological sensors.

Design principles of a robust genetic switch using antisense transcription in naturally occurring systems

249th National Meeting (2015)

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