In cellulo metrology for engineering liquid phase cellular entry

This project is inspired by the pioneering technology developed by PartitionBio (an external partner of the project) for targeted gene delivery, dubbed ‘bubbles’. The bubbles are engineered peptide-based systems that are assembled using liquid-liquid phase separation (LLPS) and enter mammalian cells by fusing with their membranes via micropinocytosis. The bubbles are unconventional gene delivery systems in both composition and mechanism of action, which differ from other cellular entry systems, both non-viral or viral. The bubbles are not structurally conserved as viruses, whose loading capacities are limited to the sizes of their genomes, and are not as elaborated architectures as DNA origami, which exhibit poor uptake. They effectively encapsulate small and large cargos such as DNA, RNA, CRISPR, or antibodies, are scalable, non-toxic and non-immunogenic, promote high payload delivery, and removal of cold chain requirements for storage and evaluations. All in all, this technology inspires liquid-phase approaches for cellular entry which hold promise for size-independent gene integration and high-content screening in engineering biology. Conversely, however, comparative or correlative data for such approaches to inform engineering rationale is lacking. This project will investigate bubble-inspired modalities in a continuum of physicochemical and biological properties to introduce benchmarking datasets for liquid-phase cellular entry – in cellulo metrology. The student will elucidate comparative metrics and parameters for membrane transfer, intracellular targeting and trafficking and will formulate the first-in-class rationale for engineering liquid-phase cellular entry.