Description:
Light Activating Genetic Tools for Rapid and Reversible Clustering of Proteins In Vivo
` Princeton Docket # 16-3266-1, 17-3279-1
Researchers in the Department of Chemical and Biological Engineering at Princeton University have developed two novel molecular tools for rapid and reversible clustering of proteins in response to light activation.
These tools will allow proteins to be clustered to form "Optodroplets", in a highly localized fashion within living cells. The clusters may also be tunable, such that they can be induced to assemble into liquid-like droplets or more solid-like gels, depending on light activation levels. It can be utilized for assembling large protein assemblies throughout the cell, or within specific sub-regions of the cell. It can also be used with lysates from cells expressing these constructs.
This technology can be used in research as a construct which could be introduced into living cells for biological studies of protein activation/inactivation, manipulation of signaling pathways, and gene expression, and also for research and industrial protein purification needs, analogous to a protein affinity tag. Proteins could be expressed in cells (e.g. bacteria or insect cells) with these light activateable protein assemblies, instead of an affinity tag such as His. The cell lysate would then be activated using blue light to induce the formation of clusters of the tagged protein. The clusters could then be centrifuged out of the solution, to high purity. The protein purification method could be made/manufactured as a small "kit" which includes a blue-light lamp, or mm-sized LEDs which would be incubated with the cell lysate to induce protein clustering.
Applications
• Manipulation of
Intracellular signaling pathway
RNA processing
Gene regulation
• Protein purification
Advantages
• Fast assembly and disassembly rate for optodrop protein
• Optodrop proteins can be clustered in highly localized fashion within cells
• Can be tuned to form liquid-like droplets or solid-like gels
Inventor
Cliff Brangwynne is an Assistant Professor in the Department of Chemical and Biological Engineering. His research focuses on using biophysical approaches to understand intracellular organization, with a particular focus on RNA/protein aggregation and phase transitions, and their impact on cell physiology and disease. Among many awards and honors Professor Brangwynne has won are a Searle Scholar Award (2012), NIH New Innovator Award (2012), Sloan Research Fellowship (2014), and an HHMI-Faculty Scholar Award (2016).
Publication
Shin Y, Berry J, Pannucci N, Haataja MP, Toettcher JE, Brangwynne CP. Spatiotemporal Control of Intracellular Phase
Transitions Using Light-Activated optoDroplets. Cell 168, 159-171, 2017.
Paci G, Lernke EA. Shining a Light on Phase Separation in the Cell. Cell 168 (1-2), 11-13. 2017
Intellectual Property & Development status
Patent protection is pending.
Princeton is currently seeking commercial partners for the further development and commercialization of this opportunity.
Contact
Laurie Tzodikov
Princeton University Office of Technology Licensing • (609) 258-7256• tzodikov@princeton.edu
Sangeeta Bafna
Princeton University Office of Technology Licensing • (609) 258-5579• sbafna@princeton.edu