CRISPRi-based circuits for genetic computation in plants

Adil Khan, Gabrielle Herring, Marina Oliva, Elliott Fourie, Jia Yuan Zhu, Benjamin Johnston, Jahnvi Pflueger, Tessa Swain, Christian Pflueger, James Peter Brook Lloyd, David Secco, Ian Small, Brendan Kidd, Ryan Lister

Research output: Working paperPreprint


Synthetic gene circuits can enable new cellular behaviours by integrating multiple input signals into customisable genetic programs. However, gene circuit development in plants has been limited by a lack of orthogonal and modular parts required for their construction. Here, we present a tool-kit of reversible CRISPRi-based gene circuits for use in plants. First, we created a range of engineered repressible promoters of different strengths and used them as integrators for the construction of NOT and NOR gates in Arabidopsis cells. Next, we determined the optimal processing system to express sgRNAs from RNA Pol II promoters to introduce NOR gate programmability and interface it with host regulatory sequences. Finally, we connected multiple NOR gates together in layered arrangements to create OR, NIMPLY, and AND logic functions. Our CRISPRi circuits are orthogonal, compact, reversible, programmable, and modular, providing a new platform for sophisticated and deliberate spatio-temporal control of gene expression in plants.
Original languageEnglish
Publication statusPublished - 2022


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