Journal Article

Design of a palette of SNAP-tag mimics of fluorescent proteins and their use as cell reporters

Chen, Xianjun, Wang, Shuo, Zhu, Linyong, Lin, Qiuning, Du, Zengmin, Hua, Yujie, Yang, Lipeng, Ge, Yihui, Li, Ningfeng, Liu, Renmei, Su, Ni, Zuo, Fangting, Bao, Chunyan, Chen, Zhengda, Jiang, Li, Zhao, Yuzheng, Hua, Xin, Bao, Bingkun, Zhang, Dasheng, Loscalzo, Joseph, Yang, Yi

2023

Overview

Naturally occurring fluorescent proteins (FPs) are the most widely used tools for tracking cellular proteins and sensing cellular events. Here, we chemically evolved the self-labeling SNAP-tag into a palette of SNAP-tag mimics of fluorescent proteins (SmFPs) that possess bright, rapidly inducible fluorescence ranging from cyan to infrared. SmFPs are integral chemical-genetic entities based on the same fluorogenic principle as FPs, i.e., induction of fluorescence of non-emitting molecular rotors by conformational locking. We demonstrate the usefulness of these SmFPs in real-time tracking of protein expression, degradation, binding interactions, trafficking, and assembly, and show that these optimally designed SmFPs outperform FPs like GFP in many important ways. We further show that the fluorescence of circularly permuted SmFPs is sensitive to the conformational changes of their fusion partners, and that these fusion partners can be used for the development of single SmFP-based genetically encoded calcium sensors for live cell imaging.

• Publisher: Springer Nature Singapore,Springer Nature B.V,Nature Publishing Group
• Subject: 631/1647 / 631/80/2373 / Biomedical and Life Sciences / Cell Biology / Cell Culture / Cell Cycle Analysis / Cell Physiology / Design / Fluorescence / Genetic engineering / Green fluorescent protein / Labeling / Life Sciences / Ligands / Protein expression / Proteins / Stem Cells