Synthetic biomolecular condensates to engineer eukaryotic cells.

The compartmentalization of specific functions into specialized organelles is a key feature of eukaryotic life. In particular, dynamic biomolecular condensates that are not membrane enclosed offer exciting opportunities for synthetic biology. In recent years, multiple approaches to generate and control condensates have been reported. Notably, multiple orthogonally translating organelles were designed that enable precise protein engineering inside living cells. Despite being built from only very few components, orthogonal translation can be engineered with subresolution precision at different places inside the same cell to create mammalian cells with multiple expanded genetic codes. This provides a pathway to engineer multiple proteins with multiple and distinct functionalities inside living eukaryotes and provides a general strategy toward spatially orthogonal enzyme engineering. [Display omitted] • Synthetic organelle formation can be controlled with small molecules or light. • Synthetic membraneless organelles enable mRNA selective genetic code expansion. • Two-dimensional phase separation creates orthogonal enzymes by subresolution spatial separation. • Film-like organelles generate cells with two expanded genetic codes. [ABSTRACT FROM AUTHOR]