Template-based copying in chemically fuelled dynamic combinatorial libraries

• Published in: Nature chemistry 2024-08, Vol.16 (8), p.1240-1249
• Main Authors: Kriebisch, Christine M. E., Burger, Ludwig, Zozulia, Oleksii, Stasi, Michele, Floroni, Alexander, Braun, Dieter, Gerland, Ulrich, Boekhoven, Job
• Format: Article
• Language: English
• Subjects: 639/638/541/960; 639/638/541/962; 639/638/904; Analytical Chemistry; Biochemistry; Chemical synthesis; Chemistry; Chemistry and Materials Science; Chemistry/Food Science; Combinatorial analysis; Combinatorial libraries; Continuous production; Copying; Coupling (molecular); Denaturation; Hybridization; Inorganic Chemistry; Oligomerization; Oligomers; Organic Chemistry; Physical Chemistry; Physical properties; Protein purification; Transcription
• ISSN: 1755-4330; 1755-4349; 1755-4349
• DOI: 10.1038/s41557-024-01570-5

One of science’s greatest challenges is determining how life can spontaneously emerge from a mixture of molecules. A complicating factor is that life and its molecules are inherently unstable—RNA and proteins are prone to hydrolysis and denaturation. For the de novo synthesis of life or to better understand its emergence at its origin, selection mechanisms are needed for unstable molecules. Here we present a chemically fuelled dynamic combinatorial library to model RNA oligomerization and deoligomerization and shine new light on selection and purification mechanisms under kinetic control. In the experiments, oligomers can only be sustained by continuous production. Hybridization is a powerful tool for selecting unstable molecules, offering feedback on oligomerization and deoligomerization rates. Moreover, we find that templation can be used to purify libraries of oligomers. In addition, template-assisted formation of oligomers within coacervate-based protocells changes its compartment’s physical properties, such as their ability to fuse. Such reciprocal coupling between oligomer production and physical properties is a key step towards synthetic life. Selection mechanisms were critical at the emergence of life and will also be important for the synthesis of life. Now, it has been shown that template-based copying controls the selection of unstable molecules in a chemically fuelled dynamic combinatorial library. Moreover, when encapsulated inside coacervate droplets, these mechanisms change the coacervate’s physical properties.