Dynamic modulation of enzyme activity by synthetic CRISPR–Cas6 endonucleases
Date
2022-04-25
Journal Title
Journal ISSN
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Publisher
Nature Chemical Biology
Abstract
In nature, dynamic interactions between enzymes play a crucial role in defining cellular metabolism. By controlling the spatial and temporal organization of these supramolecular complexes called metabolons, natural metabolism can be tuned in a highly dynamic manner. Here, we repurpose the CRISPR–Cas6 family proteins as a synthetic strategy to create dynamic metabolons by combining the ease of RNA processing and the predictability of RNA hybridization for protein assembly. By disturbing RNA–RNA networks using toehold-mediated strand displacement reactions, on-demand assembly and disassembly are achieved using both synthetic RNA triggers and mCherry messenger RNA. Both direct and ‘Turn-On’ assembly of the pathway enzymes tryptophan-2-monooxygenase and indoleacetamide hydrolase can enhance indole-3-acetic acid production by up to ninefold. Even multimeric enzymes can be assembled to improve malate production by threefold. By interfacing with endogenous mRNAs, more complex metabolons may be constructed, resulting in a self-responsive metabolic machinery capable of adapting to changing cellular demand.
Description
This article was originally published in Nature Chemical Biology. The version of record is available at: https://doi.org/10.1038/s41589-022-01005-7. This article will be embargoed until 10/25/2022.
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Citation
Mitkas, A.A., Valverde, M. & Chen, W. Dynamic modulation of enzyme activity by synthetic CRISPR–Cas6 endonucleases. Nat Chem Biol 18, 492–500 (2022). https://doi.org/10.1038/s41589-022-01005-7