Multiple designer organelles for expanded eukaryotic life
The emergence of organelles dedicated to specific cellular functions drove the evolution of more complex eukaryotic organisms. We recently created membraneless organelles inside eukaryotic cells dedicated to orthogonal translation, which opened a new path to residue-specific protein engineering using genetic code expansion. We now want to design novel organelles into eukaryotes that will internally enact the entire central dogma of molecular biology. This will supplement the complex eukaryotic cell with an additional simple and easily tailored orthogonal machinery that can also facilitate transcription and replication. This will enable us to create eukaryotes that have more than four additional expanded genetic codes, and we will explore the functional space occupied by these novel living systems. The organelles will be enhanced to process specific signals to e.g. modify RNA or degrade specific proteins. Besides these curiosity-driven goals, specific applications will allow us to road test our technology. We will directly use these approaches to advance protein engineering in eukaryotes to create proteins and artificial peptide polymers having multiple, noncanonical functionalities suitable for diverse biotechnological applications and new bioinspired materials. We will also develop organelle design into a truly universal and powerful labeling method fully compatible with eukaryotic host cell physiology that has single-residue precision and goes way beyond the state-of-the-art of any fluorescent labeling technology. The approaches will be general and truly flexible in how translation can be tailored in terms of protein, RNA and codon choice, including sense codons and type of new functionalities. Progress made in recent decades has shown that protein design and engineering can revolutionize biology. We can only imagine what can be achieved with designed functional organelles inside eukaryotic cells and how they might enable the creation of new living systems.