Optogenetic control of RNA function and metabolism using engineered light-switchable RNA-binding proteins

Renmei Liu; Jing Yang; Jing Yao; Zhou Zhao; Wei He; Ni Su; Zeyi Zhang; Chenxia Zhang; Zhuo Zhang; Haibo Cai; Linyong Zhu; Yuzheng Zhao; Shu Quan; Xianjun Chen; Yi Yang

doi:10.1038/s41587-021-01112-1

Optogenetic control of RNA function and metabolism using engineered light-switchable RNA-binding proteins

Nat Biotechnol. 2022 May;40(5):779-786. doi: 10.1038/s41587-021-01112-1. Epub 2022 Jan 3.

Authors

Renmei Liu^#^{1

2}, Jing Yang^#^{1

2}, Jing Yao^{1

2}, Zhou Zhao^{1

2}, Wei He¹, Ni Su^{1

2}, Zeyi Zhang^{1

2}, Chenxia Zhang^{1

2}, Zhuo Zhang¹, Haibo Cai¹, Linyong Zhu^{1

3}, Yuzheng Zhao^{1

2}, Shu Quan¹, Xianjun Chen^{4

5

6}, Yi Yang^{7

8

9}

Affiliations

¹ Optogenetics & Synthetic Biology Interdisciplinary Research Center, State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, China.
² School of Pharmacy, East China University of Science and Technology, Shanghai, China.
³ School of Chemistry and Molecular Engineering, East China University of Science and Technology, Shanghai, China.
⁴ Optogenetics & Synthetic Biology Interdisciplinary Research Center, State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, China. xianjunchen@ecust.edu.cn.
⁵ School of Pharmacy, East China University of Science and Technology, Shanghai, China. xianjunchen@ecust.edu.cn.
⁶ CAS Center for Excellence in Brain Science and Intelligence Technology, Chinese Academy of Sciences, Shanghai, China. xianjunchen@ecust.edu.cn.
⁷ Optogenetics & Synthetic Biology Interdisciplinary Research Center, State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, China. yiyang@ecust.edu.cn.
⁸ School of Pharmacy, East China University of Science and Technology, Shanghai, China. yiyang@ecust.edu.cn.
⁹ CAS Center for Excellence in Brain Science and Intelligence Technology, Chinese Academy of Sciences, Shanghai, China. yiyang@ecust.edu.cn.

^# Contributed equally.

PMID: 34980910
DOI: 10.1038/s41587-021-01112-1

Abstract

RNA-binding proteins (RBPs) play an essential role in regulating the function of RNAs in a cellular context, but our ability to control RBP activity in time and space is limited. Here, we describe the engineering of LicV, a photoswitchable RBP that binds to a specific RNA sequence in response to blue light irradiation. When fused to various RNA effectors, LicV allows for optogenetic control of RNA localization, splicing, translation and stability in cell culture. Furthermore, LicV-assisted CRISPR-Cas systems allow for efficient and tunable photoswitchable regulation of transcription and genomic locus labeling. These data demonstrate that the photoswitchable RBP LicV can serve as a programmable scaffold for the spatiotemporal control of synthetic RNA effectors.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

CRISPR-Cas Systems / genetics
Genomics
Optogenetics*
RNA* / genetics
RNA-Binding Proteins / genetics
RNA-Binding Proteins / metabolism

Substances

RNA-Binding Proteins
RNA