Optogenetic tool for rapid and reversible clustering of proteins

Abstract

A protein construct including a gene encoding a light-sensitive protein fused to at least one of either a low complexity sequence, an intrinsically disordered protein region (IDR), or a repeating sequence of a linker and another gene encoding a light-sensitive protein. Among the many different possibilities contemplated, the protein construct may also advantageously include cleavage tags. This protein construct may be utilized for a variety of functions, including a method for protein purification, which requires introducing the protein construct into a living cell, and inducing the formation of clusters by irradiating the construct with light. The method may also advantageously include cleaving a target protein from an IDR, and separating the clusters via centrifuge. A kit for practicing in vivo aggregation or liquid-liquid phase separation is also included, the kit including the protein construct and a light source capable of producing a wavelength that the light-sensitive protein will respond to.

What is claimed is: 
     
         1 . A method for inducing the formation of clusters or nucleation of pathological protein aggregates, comprising the steps of:
 providing a protein construct comprising:
 a first segment comprising at least one gene encoding at least one protein sensitive to at least one wavelength of light; and 
 a second segment fused to the first segment, the second segment comprising at least one intrinsically disordered protein region (IDR); and 
   inducing the protein construct to cluster by exposing the protein construct to at least one predetermined wavelength of light at a predetermined intensity for predetermined periods of time.