Organelle-Specific Nitric Oxide Detection in Living Cells via HaloTag Protein Labeling.
Nitric oxide (NO) is a membrane-permeable signaling molecule that is constantly produced, transferred, and consumed in vivo. NO participates and plays important roles in multiple biological processes. However, spatiotemporal imaging of NO in living cells is challenging. To fill the gap in currently used techniques, we exploited the versatility of HaloTag technology and synthesized a novel organelle-targetable fluorescent probe called HTDAF-2DA. We demonstrate the utility of the probe by monitoring subcellular NO dynamics. The developed strategy enables precise determination of local NO function.
Synthetic Biology and Biotechnology Laboratory, State Key Laboratory of Bioreactor Engineering, CAS Center for Excellence in Brain Science, School of Pharmacy, East China University of Science and Technology, 130 Mei Long Road, Shanghai, 200237, P. R. China.
Jianhua
J
Wang
Synthetic Biology and Biotechnology Laboratory, State Key Laboratory of Bioreactor Engineering, CAS Center for Excellence in Brain Science, School of Pharmacy, East China University of Science and Technology, 130 Mei Long Road, Shanghai, 200237, P. R. China.
Yuzheng
Y
Zhao
Shanghai Key Laboratory of Advanced Polymeric Materials, School of Materials Science and Engineering, East China University of Science and Technology, 130 Mei Long Road, Shanghai, 200237, P. R. China.
Chao
C
Wang
Synthetic Biology and Biotechnology Laboratory, State Key Laboratory of Bioreactor Engineering, CAS Center for Excellence in Brain Science, School of Pharmacy, East China University of Science and Technology, 130 Mei Long Road, Shanghai, 200237, P. R. China.
Qian
Q
Zhu
Synthetic Biology and Biotechnology Laboratory, State Key Laboratory of Bioreactor Engineering, CAS Center for Excellence in Brain Science, School of Pharmacy, East China University of Science and Technology, 130 Mei Long Road, Shanghai, 200237, P. R. China.
Zengmin
Z
Du
Shanghai Key Laboratory of Advanced Polymeric Materials, School of Materials Science and Engineering, East China University of Science and Technology, 130 Mei Long Road, Shanghai, 200237, P. R. China.
Aiguo
A
Hu
Synthetic Biology and Biotechnology Laboratory, State Key Laboratory of Bioreactor Engineering, CAS Center for Excellence in Brain Science, School of Pharmacy, East China University of Science and Technology, 130 Mei Long Road, Shanghai, 200237, P. R. China.
Yi
Y
Yang
D000478:Alkylation; D001419:Bacteria / Q000201:enzymology; D002470:Cell Survival; D002712:Chlorides / Q000737:chemistry; D005456:Fluorescent Dyes / Q000737:chemistry*; D006367:HeLa Cells; D022781:Hepatocytes; D006801:Humans; D006867:Hydrolases / Q000737:chemistry; D061986:MCF-7 Cells; D008856:Microscopy, Fluorescence; D009569:Nitric Oxide / Q000032:analysis*; D061848:Optical Imaging; D015388:Organelles / Q000737:chemistry*; D013194:Staining and Labeling
D002712:Chlorides; D005456:Fluorescent Dyes; D009569:Nitric Oxide; D006867:Hydrolases; C046137:haloalkane dehalogenase