Photogenetics technology and the method of controlling intracellular phase separation by light have been developed in recent years. Living cells can divide specific biomolecules into different compartments, so that biochemical reactions can be carried out relatively independently, efficiently and orderly. One compartment is covered with biofilm. In addition, there are a large number of membranous compartments, including P particles of reproductive cells of Caenorhabditis elegans, stress particles involved in stress response, p62 particles involved in self spitting activities, etc. they are called "biomolecular aggregates". Because these compartments are membrane-free, they can exchange molecules quickly with the outside and play an important role in intracellular signal transduction. Therefore, the formation process, physical properties, chemical properties and cell function of biomolecular aggregates have become a research hotspot in biology and its cross fields.

Optogenetic Tools for Cell Phase Separation Research

Most bio molecular aggregates, such as nucleoli and stress particles, are formed by "liquid-liquid phase separation". This interaction is usually driven by the polymerization of intrinsic disordered regions (IDR) of proteins. The precise manipulation of intracellular phase separation process using optogenetic tools will help researchers deepen their understanding of the specific mechanism and regulation process of intracellular phase separation.

Achievable Missions at Creative BioMart

Creative BioMart has mastered and developed a variety of optical switching systems for cell phase separation. We have the ability to provide the construction services of cell lines, expression systems and model organisms.

Reversible control of intracellular phase separation—blue light

IDR-CRY2 fusion protein induced the formation of CRY2 polymer under blue light. This principle has been used to construct an optical control system to control the phase separation process in cells. This method can realize the reversible control of intracellular phase separation and activate the phase separation process locally.

To study the process of phase separation in cells and its effect on chromosomes—blue light

The system constructed with AsLOV2 switch can be used to study the phase separation process in cells and its effect on chromosomes. The system was constructed by improved light induced dimer (iLID) and its homologous binding protein sspB (stringent stavation protein B) based on aslov2. Through the combination of iLID and sspB under blue light, a rapid response and quantifiable system under light was used to study the initial threshold protein concentration and transformation mechanism of intracellular phase separation.

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Reference

1. Shin Y, Berry J, Pannucci N, Haataja MP, Toettcher JE, Brangwynne CP. Spatiotemporal Control of Intracellular Phase Transitions Using Light-Activated optoDroplets. Cell. 2017 Jan 12;168(1-2):159-171.e14.

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