2011/10/18 by Baojun Wang, R.I. Kitney, Nicolas Joly +1 · 1 citation
Biochemistry, Genetics and Molecular Biology · #Gene Regulatory Network Analysis #Bacterial Genetics and Biotechnology #CRISPR and Genetic Engineering
paper · pdf · doi:10.1038/ncomms1516
openalex publication_date 2011/10/18 · openalex created_date 2025/10/10 · openalex updated_date 2026/08/02
Modular and orthogonal genetic logic gates are essential for building robust biologically based digital devices to customize cell signalling in synthetic biology. Here we constructed an orthogonal AND gate in Escherichia coli using a novel hetero-regulation module from Pseudomonas syringae. The device comprises two co-activating genes hrpR and hrpS controlled by separate promoter inputs, and a σ54-dependent hrpL promoter driving the output. The hrpL promoter is activated only when both genes are expressed, generating digital-like AND integration behaviour. The AND gate is demonstrated to be modular by applying new regulated promoters to the inputs, and connecting the output to a NOT gate module to produce a combinatorial NAND gate. The circuits were assembled using a parts-based engineering approach of quantitative characterization, modelling, followed by construction and testing. The results show that new genetic logic devices can be engineered predictably from novel native orthogonal biological control elements using quantitatively in-context characterized parts. Biological digital sensors require the fabrication of modular genetic logic gates. Using thePseudomonas syringae hrpsystem, Wang and colleagues generate AND, NOT and NAND gates, demonstrating the ability to engineer a modular system from biological elements.