PT  - JOURNAL ARTICLE
AU  - Hideki Nakamura
AU  - Albert A. Lee
AU  - Ali Sobhi Afshar
AU  - Shigeki Watanabe
AU  - Elmer Rho
AU  - Shiva Razavi
AU  - Allison Suarez
AU  - Yu-Chun Lin
AU  - Makoto Tanigawa
AU  - Brian Huang
AU  - Robert DeRose
AU  - Diana Bobb
AU  - William Hong
AU  - Sandra B. Gabelli
AU  - John Goutsias
AU  - Takanari Inoue
TI  - Intracellular production of hydrogels and synthetic RNA granules by multivalent enhancers
AID  - 10.1101/117572
DP  - 2017 Jan 01
TA  - bioRxiv
PG  - 117572
4099  - http://biorxiv.org/content/early/2017/03/16/117572.short
4100  - http://biorxiv.org/content/early/2017/03/16/117572.full
AB  - Non-membrane bound, hydrogel-like entities, such as RNA granules, nucleate essential cellular functions through their unique physico-chemical properties. However, these intracellular hydrogels have not been as extensively studied as their extracellular counterparts, primarily due to technical challenges in probing these materials in situ. Here, by taking advantage of a chemically inducible dimerization paradigm, we developed iPOLYMER, a strategy for rapid induction of protein-based hydrogels inside living cells. A series of biochemical and biophysical characterizations, in conjunction with computational modeling, revealed that the polymer network formed in the cytosol resembles a physiological hydrogel-like entity that behaves as a size-dependent molecular sieve. We studied several properties of the gel and functionalized it with RNA binding motifs that sequester polyadenine-containing nucleotides to synthetically mimic RNA granules. Therefore, we here demonstrate that iPOLYMER presents a unique and powerful approach to synthetically reconstitute hydrogel-like structures including RNA granules in intact cells.