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Peking 2018
Synthetic Phase separation-based Organelle Platform (SPOT) Abstract
(一) Background and motivation
Ever since the beginning of life, compartment has been playing a crucial rule in biological systems. The famous Miller-Urey experiment shows that inorganic molecules can transform into organic substances under extreme conditions, for example lightening. However, homogeneously distributed organic matters are not enough for life to emerge. It is almost impossible that all conditions are proper in the entire primordial soup.
That (pause) is where the compartment comes in.
Only after coacervate droplet forms and organic molecules condense inside, a completely different environment can be attained within, thus enabling the emergence of bio-macromolecules, or in other word, making life possible.
In cells, compartmentalization is mainly achieved be all sorts of organelles, for instance, mitochondrion, chloroplast, lysosome etc. They take up three major roles: A, B, C
Intuitively, for a organelle to sustain a stable compartment, it seems necessary to require a material boundary, more precisely, a membrane. Membrane-bound organelles are indeed common and stable, but from the perspective of synthesis, it is way too complicated. However, there are also non-membrane-bound organelles, for instance, stress granule, P granule and nucleolus. More importantly, their formation is guided by simple physical principals.Membrane-less organelles and phase separation. Then came the question that how can we synthase membraneless organelles.
(二) Principles and design
There are a large number of phase separation phenomena in cells, summarizing which, we put forward a principle that interaction and multivalence are two preconditions of phase separation in cells. Based on this principle, we use SUMO/SIM, FKBP/Frb, etc. as interactional modules to provide diverse inducing of the condensation while we fuse homo-oligometric tags (HOTags) to introduce multivalence. We named our system SPOT(Synthetic Phase separation-based Organelle Platform) because it can form granules( we can see fluorescent spots in yeast under microscope) in yeast.
(三) SPOT construction and verification
We try diverse interactional modules to construct the synthetic organelles and then modeled our system according to the theory of phase separation. As this model predicts, different promoters alter the features and kinetics of our systems, which were well validated by experiments
(四) Functions of synthetic organelles
We verified the feasibility of several potential functions theoretically and experimentally, including reaction crucible and sensor, etc. In the future, by replacing functional modules with other parts, this system would conduct functions not included in the current project.