Chlorophyll-Based Optogenetics to Control Membraneless Organelles

Manjia Li; Byung Min Park; Zhaoxia Li; Weiqi Huang; Fei Sun

doi:10.1007/978-1-0716-4047-0_15

Chlorophyll-Based Optogenetics to Control Membraneless Organelles

Methods Mol Biol. 2025:2840:201-216. doi: 10.1007/978-1-0716-4047-0_15.

Authors

Manjia Li¹, Byung Min Park^{1

2}, Zhaoxia Li³, Weiqi Huang⁴, Fei Sun^{5

6

7}

Affiliations

¹ Department of Chemical and Biological Engineering, The Hong Kong University of Science and Technology, Hong Kong, China.
² Department of Imaging Physics, Faculty of Applied Sciences, Delft University of Technology, Delft, the Netherlands.
³ College of Marine and Biological Engineering, Yancheng Institute of Technology, Yancheng, Jiangsu, China.
⁴ College of Computer and Information Sciences, Fujian Agriculture and Forestry University, Fuzhou, Fujian, China.
⁵ Department of Chemical and Biological Engineering, The Hong Kong University of Science and Technology, Hong Kong, China. [email protected].
⁶ Greater Bay Biomedical InnoCenter, Shenzhen Bay Laboratory, Shenzhen, China. [email protected].
⁷ Research Institute of Tsinghua, Pearl River Delta, Guangzhou, China. [email protected].

PMID: 39724354
DOI: 10.1007/978-1-0716-4047-0_15

Abstract

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.

Keywords: Chlorophyll; Membraneless organelle; Optogenetics; Photo-responsive protein; Protein phase transition.

MeSH terms

Chlorophyll* / metabolism
Humans
Light
Optogenetics* / methods
Organelles* / metabolism
Singlet Oxygen / metabolism

Substances

Chlorophyll
Singlet Oxygen