[3S04m] 11. The Society for Biotechnology - Part 1

11:30 AM - 11:45 AM JST(2:30 AM - 2:45 AM UTC)

[3SBT-06] Ribosome biogenesis in vitro

○Wataru Aoki¹ (1. The University of Osaka (Japan))

Keywords:

Ribosome biogenesis,Artificial cell

The minimal mechanism for the self-replication of life is the autonomous central dogma. This is defined as a system that sustains self-replication by executing transcription, translation, DNA replication, and ribosome biogenesis based on its own encoded genetic information. To fully understand the self-replication process, it is essential to reconstitute these four fundamental processes in vitro and reproduce this autonomous central dogma in a cell-free environment. While the reconstitution of transcription, translation, and DNA replication was achieved nearly sixty years ago, the in vitro reconstitution of ribosome biogenesis has remained elusive due to its extreme complexity. Ribosome biogenesis is pivotal in the self-replication of life. In Escherichia coli, three ribosomal RNAs and 54 ribosomal proteins are synthesized and subjected to cooperative hierarchical assembly facilitated by numerous accessory factors. Realizing ribosome biogenesis in vitro is an essential goal to understand the self-replication of life and creating artificial cells; however, this has not been realized because of its complexity. We have reported the first successful ribosome biogenesis in vitro. Specifically, we have developed several key technologies: a methodology for the precise control of gene expression levels in vitro, mass spectrometry techniques for the comprehensive quantification of ribosomal components, and a highly specific and sensitive method for detecting only nascent ribosomes (PLoS ONE, 2018; PLoS ONE, 2020). The reporter assay allowed for combinatorial and iterative exploration of reaction conditions for ribosome biogenesis, leading to the simultaneous, autonomous synthesis of both small and large subunits of ribosomes in vitro through transcription, translation, processing, and assembly in a single reaction space (Nature Communications, 2025). Our achievement represents a critical step toward revealing the fundamental principles underlying the self-replication of life and creating artificial cells.