Higher-order structure of DNA determines its positioning in cell-size droplets under crowded conditions

Background It is becoming clearer that living cells use water/water (w/w) phase separation to form membraneless organelles that exhibit various important biological functions. Currently, it is believed that the specific localization of biomacromolecules, including DNA, RNA and proteins in w/w microdroplets is closely related to their bio-activity. Despite the importance of this possible role of micro segregation, our understanding of the underlying physico-chemical mechanism is still unrefined. Further research to unveil the underlying mechanism of the localization of macromolecules in relation to their steric conformation in w/w microdroplets is needed. Principal findings Single-DNA observation of genome-size DNA (T4 GT7 bacteriophage DNA; 166kbp) by fluorescence microscopy revealed that DNAs are spontaneously incorporated into w/w microdroplets generated in a binary aqueous polymer solution with polyethylene glycol (PEG) and dextran (DEX). Interestingly, DNAs with elongated coil and shrunken conformations exhibit Brownian fluctuation inside the droplet. On the other hand, tightly packed compact globules, as well as assemblies of multiple condensed DNAs, tend to be located near the interface in the droplet. Conclusion and significance The specific localization of DNA molecules depending on their higher-order structure occurs in w/w microdroplet phase-separation solution under a binary aqueous polymer solution. Such an aqueous solution with polymers mimics the crowded conditions in living cells, where aqueous macromolecules exist at a level of 30–40 weight %. The specific positioning of DNA depending on its higher-order structure in w/w microdroplets is expected to provide novel insights into the mechanism and function of membraneless organelles and micro-segregated particles in living cells.

Unravelling a Novel Sol-Gel Transition Mechanism in Polymer Self-Assemblies: An Order-Order Transition Based on Specific Molecular Interactions Between Hydrophilic and Hydrophobic Polymer Blocks

Using a wide range of state-of-the art analytical techniques and molecular dynamics simulation, a novel mechanism for macromolecular interactions are described. Distinct interactions between the hydrophilic and hydrophobic blocks in amphiphilic triblock copolymers lead to an order-order transition from spherical micelles to worm-like micelles upon cooling the aqueous polymer solutions below room temperature. Macroscopically, this this leads to reversible gelation. This novel mechanism represent a novel building block to better understand polymer self-assembly.<br>

Unravelling a Novel Sol-Gel Transition Mechanism in Polymer Self-Assemblies: An Order-Order Transition Based on Specific Molecular Interactions Between Hydrophilic and Hydrophobic Polymer Blocks

Using a wide range of state-of-the art analytical techniques and molecular dynamics simulation, a novel mechanism for macromolecular interactions are described. Distinct interactions between the hydrophilic and hydrophobic blocks in amphiphilic triblock copolymers lead to an order-order transition from spherical micelles to worm-like micelles upon cooling the aqueous polymer solutions below room temperature. Macroscopically, this this leads to reversible gelation. This novel mechanism represent a novel building block to better understand polymer self-assembly.<br>

An Inverse Thermogelling Bioink Based on an ABA Type Poly(2-Oxazoline) Amphiphile

Hydrogels are key components in several biomedical research areas such as biofabrication. Here, a novel ABA-type triblock copolymer is introduced that undergoes a inverse thermogelation, i.e. it forms a hydrogel about cooling aqueous polymer solutions. The macroporous hydrogel was rheologically investigate in detail and used for 3D printing using an extrusion based printer. Preliminary experiment show very good cytocompatibility of NIH 3T3 cells also after printing and the possibility to combine the novel material with other hydrogel forming materials such as alginate.<br>