Functional design of ribosomes with mutant ribosomal RNA (rRNA) could expand opportunities for understanding molecular translation, building cells from the bottom-up, and engineering ribosomes with altered capabilities. However, such efforts have been hampered by cell viability constraints, an enormous combinatorial sequence space, and limitations on large-scale, 3D design of RNA structures and functions. To address these challenges, we developed an integrated community science and experimental screening approach for rational design of ribosomes. This approach couples Eterna, an online video game that crowdsources RNA sequence design to community scientists in the form of puzzles, with in vitro ribosome synthesis, assembly, and translation in multiple design-build-test-learn cycles. We applied our framework to discover mutant rRNA sequences that improve protein synthesis in vitro and cell growth in vivo, relative to wild type ribosomes, under diverse environmental conditions. This work provides new insights into ribosome rRNA sequence-function relationships, with implications for synthetic biology.

中文翻译：

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社区科学设计了具有有益表型的核糖体

具有突变核糖体 RNA (rRNA) 的核糖体的功能设计可以扩大理解分子翻译、自下而上构建细胞和设计具有改变能力的核糖体的机会。然而，这些努力受到细胞活力的限制、巨大的组合序列空间以及对 RNA 结构和功能的大规模 3D 设计的限制。为了应对这些挑战，我们开发了一种综合的社区科学和实验筛选方法来合理设计核糖体。这种方法将 Eterna（一种以拼图形式向社区科学家众包 RNA 序列设计的在线视频游戏）与体外核糖体合成、组装和翻译结合在多个设计-构建-测试-学习循环中。我们应用我们的框架来发现在不同环境条件下相对于野生型核糖体改善体外蛋白质合成和体内细胞生长的突变 rRNA 序列。这项工作提供了对核糖体 rRNA 序列-功能关系的新见解，对合成生物学有影响。