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Award Abstract #1253035

CAREER: Non-Equilibrium RNA/Protein Liquids and Intracellular Phase Transitions

NSF Org: PHY
Division Of Physics
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Initial Amendment Date: September 5, 2013
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Latest Amendment Date: August 13, 2015
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Award Number: 1253035
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Award Instrument: Continuing grant
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Program Manager: Krastan B. Blagoev
PHY Division Of Physics
MPS Direct For Mathematical & Physical Scien
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Start Date: September 15, 2013
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End Date: August 31, 2018 (Estimated)
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Awarded Amount to Date: $420,000.00
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Investigator(s): Clifford Brangwynne cbrangwy@princeton.edu (Principal Investigator)
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Sponsor: Princeton University
Off. of Research & Proj. Admin.
Princeton, NJ 08544-2020 (609)258-3090
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NSF Program(s): PHYSICS OF LIVING SYSTEMS,
Cellular Dynamics and Function
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Program Reference Code(s): 7237, 7246, 8007, 9183, 1045
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Program Element Code(s): 7246, 1114

ABSTRACT

In this project, the PI will focus on elucidating two important aspects of RNP liquids: 1) Their assembly from soluble components, and 2) Their material properties, which strongly impact their biological function. The PI will study the complexity of living cells by first taking a bottom-up approach, utilizing an in vitro system of purified components. This will be complemented by utilizing advanced microscopy and image analysis techniques, and novel molecular dynamics measurements. These combined in vitro/in vivo studies will allow the PI to understand the full complexity of RNP liquids, and test the hypothesis that the assembly and properties of these droplets are tuned by ATP-dependent biological activity. The broader impacts of this proposal include educating the next generation of biophysicists by providing laboratory research training at the high school, undergraduate, and graduate levels; at the undergraduate level, laboratory training for senior thesis research will play a key role. In the classroom, the PI will develop new courses on Quantitative Biology and Soft Living Matter. This teaching is part of the PI's broader effort to build the infrastructure for growing Princeton's nascent Bioengineering community, and couple it to the greater Biophysics community on campus and beyond. At the K-12 level, the PI has developed an outreach partnership with the Trenton public schools, which focuses on inspiring under-represented groups of 8th grade students from Kilmer Elementary School. The PI will visit the classroom for hands-on presentations on Soft Living Matter, and these students will visit the PI's laboratory. To evaluate the efficacy of this outreach, quantitative evaluation tools based on the Test of Science Related Attitudes (TOSRA) will be used, which the PI will implement and analyze together with a science education researcher.

This project is being jointly supported by the Physics of Living Systems program in the Division of Physics and the Cellular Dynamics and Function Program in the Division of Molecular and Cellular Biosciences.


PUBLICATIONS PRODUCED AS A RESULT OF THIS RESEARCH

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L. Zhu, C.P. Brangwynne. "Nuclear Bodies: The emerging biophysics of nucleoplasmic phases," Current Opinion in Cell Biology, v.34, 2015, p. 23. 

S. Elbaum-Garfinkle, Y. Kim, C. Szczepaniak, C. Eckmann, S. Myong, C.P Brangwynne. "The disordered P granule protein LAF-1 drives phase separation into droplets with tunable viscosity and dynamics," Proceedings of the National Academy of Science USA, v.112, 2015, p. 7189. 

S.C. Weber, C.P. Brangwynne. "Inverse Size Scaling of the Nucleolus by a Concentration-Dependent Phase Transition," Current Biology, v.25, 2015, p. 641. 

W. Gilpin, S. Uppaluri. "Worms under pressure: bulk mechanical properties of C.elegans are independent of the cuticle," Biophysical Journal, v.108, 2015, p. 1887. 

C.P. Brangwynne, P.S. Tompa, R.V. Pappu. "Phase transitions and the polymer physics of intracellular organization," Nature Physics, v.11, 2015, p. 899. 

H. Zhang, S. Elbaum-Garfinkle, E. Langdon, N. Taylor, P. Occhipinti, A. Bridges, C.P. Brangwynne, A.S. Gladfelter. "RNA controls PolyQ protein phase separation in a sequence specific manner," Molecular Cell, v.60, 2015, p. 220. 

J. Berry, S.C. Weber, N. Vaidya, M. Haataja, C.P. Brangwynne. "RNA Transcription Modulates Phase Transition-Driven Nuclear Body Assembly," Proceedings of the National Academy of Science USA, v.112, 2015, p. E5237. 

J. Bosse, I.B. Hogue, M. Feric, S.Y. Thiberge, B. Sodeik, C.P. Brangwynne, L.W. Enquist. "Remodeling nuclear architecture allows efficient transport of herpesvirus capsids by diffusion," Proceedings of the National Academy of Sciences USA, v.112, 2015, p. E5725. 

M. Feric, C.Broedersz, C.P. Brangwynne. "Soft viscoelastic properties of nuclear actin age oocytes due to gravitational creep," Scientific Reports, 2015. 

Feric M, Vaidya N, Harmon TS, Mitrea DM, Zhu L, Richardson TM, Kriwacki RW, Pappu RV, Brangwynne CP. "Coexisting liquid phases underlie nucleolar sub-compartments," Cell, 2016. 


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