As promised, we run the town and gown 10k! In the photos are group members @rogerrubiosanchez.bsky.social Juliette Bucci, Brian Ng, Cathrine Fan, Sebastian Krauss and Lorenzo (the old one). Thanks to all who supported us with donations to @mdukcharity.bsky.social

Finally, we explore the effect on phase behaviour of changing annealing protocols, demonstrating that multicomponent DNA condensates equilibrate veeeeeeeeeeery slowly. Something to bear in mind if you plan to work with similar systems 5/n

Simulations by Dino Osmanovic @francolab.bsky.social, based on Flory-Huggins, map well onto the experimental trends and allowed us to establish a link between the experimental order parameter and the F-H interaction parameter. Experimental (top) and simulated (bottom) snapshots below 4/n

We can control phase behaviour by changing linker and nanostars concentrations, without having to re-design nanostructures. This is rather convenient, and allowed us to map a large phase diagram and identify an order parameter controlling the transition between 1- and 2-phase condensates 3/n

Condensates assemble from two populations of tetravalent DNA nanostars, and three types of divalent linkers that mediate nanostar-nanostar interactions, either within the same nanostar population or across populations 2/n

Finally, we show that condensation is thermally reversible. The MLOs melt upon heating and re-assemble upon cooling, releasing and re-capturing GFP in the process 8/n

The nanostar designs are modular and allow for embedding of protein binding aptamers. We show that GFP can be selectively captured by the MLOs when a GFP-binding aptamer is included 7/n

We can express two non-interacting nanostars within the same cell, creating two orthogonal MLOs. This is only possible thanks to the selectivity of base pairing. Also here, most cells will have both MLOs, located at or near the poles 6/n

MLOs are located at the cell poles but small RNA clusters may appear near the centre before moving towards the poles. MLOs also tend to appear in the central section as cells approach division. The new MLOs may end up in one of the daughter cells, or be split between both 5/n

We express RNA “nanostars” interacting with kissing loops (KL) in E. coli and show that these form MLOs located at the poles of the cell. Expression is inducible and very efficient. Making the nanostars non-sticky stops condensation, indicating specificity 4/n