Determination of Intracellular Condensates Based on Optogenetic Reconstitution

Many membraneless organelles are highly dynamic structures formed by liquid-liquid phase separation (LLPS) processes. However, little is known about the biophysical mechanisms and functional consequences of these myriad dynamic changes. To study how spatial and temporal regulation affects organelle function, it is necessary to induce phase separation at the desired temporal or spatial location within the cell. Recently developed optogenetic methods are well suited for such "control-freak" experiments, where light can be sensitively delivered and focused with micron-level spatial precision, allowing selective activation of proteins at specific times or locations within cells or tissues.

Fig. 1. Optogenetic reconstitution for determining the form and function of membraneless organelles. (Dine E, et al., 2018)

Customized Services

Phase separation arises from multiple weak interactions occurring between different biomolecules, making it difficult to eliminate without counteracting effects and difficult to induce with typical biological tools. CD BioSciences offers optogenetic methods to induce LLPS at the desired temporal or spatial location in the cell.

We drastically change the concentration of photoactivated proteins by varying the intensity of blue light or the frequency of light pulses, which can quantitatively characterize their phase separation. In addition, we provide a variety of heterodimer formation tools from optogenetics and chemical biology to design a series of photoswitchable dimmers to construct controlled aggregation systems in which multivalency and interaction strength can be quantitatively controlled.

We offer comprehensive optogenetic phase separation services allowing researchers to acutely control the timing, location and composition of membrane-free organelle formation, and can even be used to induce the formation of different phases. Our services help dissect how each candidate mechanism contributes to the regulation of those cellular processes for which membraneless organelles are observed. We aim to help our clients complete the biophysical picture of how intracellular protein phase separation is controlled in vivo. Our optogenetic approach is primarily used for the following:

• Studying the morphology of intracellular biomolecular condensates.
• Determining the function of intracellular biomolecular condensates.

Advantages of Our Services

• Enable selective activation of proteins at specific times or locations within cells or tissues.
• Be highly specific.
• Can easily adjust the intensity or schedule of light transmission.
• Can be used to precisely and independently control the specific biochemical properties, such as the concentration of active proteins or even the duration of complex formation between the photosensitive protein and its binding partner.
• Determing not only the mode of formation of membrane-free organelles, but also includes a description of their function.

CD BioSciences has been focusing on how optogenetic reconstruction facilitates the production of membraneless organelles. Our experts are also trying to use optogenetic tools to further understand the biophysics behind the phase transition and the biochemical parameters it regulates. We aim to help you analyze the biophysical processes of intracellular biomolecular condensates through optical gate tools that control protein phases. If you have any special requirements for our services, please feel free to contact us. We are looking forward to working together with your attractive projects.