A general fluorescence off/on strategy for fluorogenic probes: Steric repulsion-induced twisted intramolecular charge transfer (sr-TICT)

  • Sci Adv. 2024 Feb 16;10(7):eadi8847. doi: 10.1126/sciadv.adi8847.
Kenjiro Hanaoka  1 ,  Takayuki Ikeno  2 ,  Shimpei Iwaki  2 ,  Sayaka Deguchi  3 ,  Kazuo Takayama  3 ,  Hiroyuki Mizuguchi  3  4  5 ,  Fumiya Tao  6 ,  Nobuhiko Kojima  6 ,  Hisashi Ohno  1 ,  Eita Sasaki  1 ,  Toru Komatsu  2 ,  Tasuku Ueno  2 ,  Kazuya Maeda  2 ,  Hiroyuki Kusuhara  2 ,  Yasuteru Urano  2  7
Affiliations
  • 1. Graduate School of Pharmaceutical Sciences, Keio University, 1-5-30 Shibakoen, Minoto-ku, Tokyo 105-8512, Japan.
  • 2. Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.
  • 3. Laboratory of Biochemistry and Molecular Biology, Graduate School of Pharmaceutical Sciences, Osaka University, 1-6 Yamadaoka, Suita, Osaka 565-0871, Japan.
  • 4. Integrated Frontier Research for Medical Science Division, Institute for Open and Transdisciplinary Research Initiatives, Osaka University, Osaka 565-0871, Japan.
  • 5. National Institutes of Biomedical Innovation, Health and Nutrition, Osaka 567-0085, Japan.
  • 6. Department of Life and Environmental System Science, Graduate School of Nanobioscience, Yokohama City University, 22-2 Seto, Kanazawa-ku, Yokohama 236-0027, Japan.
  • 7. Graduate School of Medicine, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.
PMID: 38363840 DOI: 10.1126/sciadv.adi8847
Abstract

Various control strategies are available for building fluorogenic probes to visualize biological events in terms of a fluorescence change. Here, we performed the time-dependent density functional theory (TD-DFT) computational analysis of the twisted intramolecular charge transfer (TICT) process in rhodamine dyes. On the basis of the results, we designed and synthesized a series of rhodamine dyes and established a fluorescence quenching strategy that we call steric repulsion-induced TICT (sr-TICT), in which the fluorescence quenching process is greatly accelerated by simple intramolecular twisting. As proof of concept of this design strategy, we used it to develop a fluorogenic probe, 2-Me PeER (pentyloxyethylrhodamine), for the N-dealkylation activity of CYP3A4. We applied 2-Me PeER for CYP3A4 activity-based fluorescence-activated cell sorting (FACS), providing access to homogeneous, highly functional human-induced pluripotent stem cell (hiPSC)-derived hepatocytes and intestinal epithelial cells. Our results suggest that sr-TICT represents a general fluorescence control method for fluorogenic probes.

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