RiboLife aimed to explore RNA-based genetic storage and replication, leading to significant advancements in synthetic RNA biology and RNA-DNA hybrid systems. Throughout the project, research was structured around three main objectives:
1. Development of artificial self-replicating RNA scaffolds
2. Engineering of self-replicating RNA nanostructures
3. Transplantation of essential cellular functions onto synthetic RNA genomes
A breakthrough of the project was the first successful replication of ribosomal RNAs (23S, 16S, and 5S, including tethered constructs) in a DNA-free system. This result demonstrates that RNA-based genetic replication can sustain core cellular functions, opening new avenues in synthetic ribosome engineering. Additionally, we identified critical bacterial host factors involved in regulating phage RNA replicases, which enabled the first complete in vitro reconstitution of the MS2 replicase complex.
Another major achievement was the development of novel self-replicating RNA scaffolds derived from bacteriophage MS2. These engineered RNAs were designed to carry functional genetic elements, including ribozymes and protein-coding genes, demonstrating their potential as synthetic gene expression platforms. Towards the later stages of the project, we successfully achieved partial intracellular RNA replication, showing that essential genes can be transplanted onto RNA-based genetic carriers, which represents an important step toward RNA-based genome engineering.
An unexpected yet highly promising result emerged when we explored other strategies to create RNA-DNA hybrid systems. We discovered that bacterial retrons can be harnessed to synthesize functional single-stranded DNA aptamers inside cells from RNA templates, a finding that holds significant potential for DNA-based therapeutics, gene silencing, and programmable nucleic acid synthesis.
Taken together, these findings significantly advance the field of synthetic RNA biology by establishing a new platform for RNA-based information storage, gene evolution, and synthetic gene expression. The ability to replicate ribosomal RNA without DNA intermediates provides a framework for designing artificial translation systems, while the retron-based RNA-DNA hybrid approach offers novel tools for intracellular genetic programming. We have published our main findings in peer-reviewed publications in high-impact journals. This includes a recent systematic review on synthetic self-replicating RNAs and their applications. Remaining key challenges that require further research concerns in particular the long-term stability of srRNAs inside living cells. Ongoing experiments aim to build on our discoveries, with potential applications in biotechnology, synthetic biology, and molecular genetics. The results of RiboLife not only provide new insights into the potential of RNA-based genetic systems but also lay the groundwork for future applications in biotechnology, synthetic biology, and molecular therapeutics.
