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Regulating Nucleic Acid Catalysis Using Active Droplets
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DOI：10.1002/anie.202412534.png)
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Cells use transient membraneless organelles to regulate biological reaction networks. For example, stress granules selectively store mRNA to downregulate protein expression in response to heat or oxidative stress. Models mimicking this active behavior should be established to better understand in vivo regulation involving compartmentalization. Here we use active, complex coacervate droplets as a model for membraneless organelles to spatiotemporally control the activity of a catalytic DNA (DNAzyme). Upon partitioning into these peptide-RNA droplets, the DNAzyme unfolds and loses its ability to catalyze the cleavage of a nucleic acid strand. We can transiently pause the DNAzyme activity upon inducing droplet formation with fuel. After fuel consumption, the DNAzyme activity autonomously restarts. We envision this system could be used to up and downregulate multiple reactions in a network, helping understand the complexity of a cell's pathways. By creating a network where the DNAzyme could reciprocally regulate the droplet properties, we would have a powerful tool for engineering synthetic cells. In this study, we use active, fuel-dependent complex coacervate droplets as a model for membraneless organelles and spatiotemporally control the catalytic activity of the 17E DNAzyme. Partitioning the biocatalyst into the peptide-RNA droplets results in unfolding and activity loss. The catalytic activity is recovered after droplets transiently dissolve when fuel is used up, mimicking they dynamic regulation of biocatalytic pathways in membraneless organelles. image
Keyword:
DNAzyme
Artificial Organelles
Transient catalysis
Coacervates
Synthetic Biology
期刊
IF:
16.9
论文数:
5.7W
被引数:
53.0W
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