To ensure continuance, animal species must make totipotent embryos for the next generation. In
sexually reproducing animals, this is achieved by the germline. Germline cells contain a specialized
non-membrane-bound compartment, containing proteins and RNAs, called Nuage. Mutations in
nuage proteins are associated with loss of germ cell fate and infertility. However, the molecular
mechanisms of nuage function remain unclear. A major bottleneck towards addressing this question
has been the inability to isolate nuage from in vivo sources. Dilutions that accompany purification
strategies led to dissolution of liquid-like nuage. We have shown that nuage-like phases can be
reconstituted in vitro from purified components, thereby providing a way around the long-standing
bottleneck in addressing the molecular mechanisms of nuage function. We will investigate the
biochemical environment within the nuage phase and its interactions with the surroundings in C.
elegans using a combination of live cell microscopy, in vitro reconstitution, biochemistry, and
structural biology approaches. In the long-term, our study on the foundations of a germ cell-specific
phenomenon may lay the groundwork for understanding problems with fertility and elucidate ways to
address them.