Decoding nucleic acid contributions to phase separation and ordering in biomolecular condensates

Liquid-liquid phase separation (LLPS) is being increasingly recognized as a major organizational principle for proteins and nucleic acids (NAs) in cells, as well as a promising strategy for synthetic biology and biomedical applications. Extensive work has explored the role of protein sequence and properties in the regulation of LLPS. Despite some relevant exceptions, the role of nucleic acids has been much less explored. Here, to fill this gap, we focus on model systems made of oligonucleotides with tuned lengths and degree of hybridization, mixed with a moderately charged, disordered peptide. Combining multiple length scales through experiments and molecular dynamics simulations, we unravel the distinct effects of the properties of NAs on the phase behavior, and we propose a metric for the stability of biocondensates. We also characterize the conditions for the onset of liquid crystalline order in the droplets, and we show that it is associated with a dramatic slowing down of the dynamics of oligonucleotides, while peptides retain high mobility. Our results can be generalized to natural and non-natural NAs of arbitrary structure, providing a guide for the design of synthetic NA-containing coacervates.