Chlorophyll-Based Optogenetics to Control Membraneless Organelles
Manjia Li (The Hong Kong University of Science and Technology)
Byung Min Park (The Hong Kong University of Science and Technology, TU Delft - ImPhys/Maresca group)
Zhaoxia Li (Yancheng Institute of Technology)
Weiqi Huang (Fujian Agriculture and Forestry University)
Fei Sun (Research Institute of Tsinghua, The Hong Kong University of Science and Technology, Shenzhen Bay Laboratory)
More Info
expand_more
Other than for strictly personal use, it is not permitted to download, forward or distribute the text or part of it, without the consent of the author(s) and/or copyright holder(s), unless the work is under an open content license such as Creative Commons.
Abstract
Protocol.
Membraneless organelles (MLOs) formed via protein phase separation have garnered significant attention recently due to their relevance to cellular physiology and pathology. However, there is a lack of tools available to study their behavior and control their bioactivity in complex biological systems. This chapter describes a new optogenetic tool based on water-soluble chlorophyll protein (WSCP), a red light-induced singlet oxygen-generating protein, to control synthetic MLOs. Upon exposure to red light, WSCP generates singlet oxygen, which triggers the crosslinking of the proteins in the MLOs, resulting in their liquid-to-solid phase transition. The effective delivery of chlorophylls enables the successful reconstitution of WSCP in living cells, thus offering a potential approach to biological regulation at the subcellular level.