Development of background-free tame fluorescent probes for intracellular live cell imaging.
Fluorescence labelling of an intracellular biomolecule in native living cells is a powerful strategy to achieve in-depth understanding of the biomolecule's roles and functions. Besides being nontoxic and specific, desirable labelling probes should be highly cell permeable without nonspecific interactions with other cellular components to warrant high signal-to-noise ratio. While it is critical, rational design for such probes is tricky. Here we report the first predictive model for cell permeable background-free probe development through optimized lipophilicity, water solubility and charged van der Waals surface area. The model was developed by utilizing high-throughput screening in combination with cheminformatics. We demonstrate its reliability by developing CO-1 and AzG-1, a cyclooctyne- and azide-containing BODIPY probe, respectively, which specifically label intracellular target organelles and engineered proteins with minimum background. The results provide an efficient strategy for development of background-free probes, referred to as 'tame' probes, and novel tools for live cell intracellular imaging.
Department of Chemistry &Med Chem Program, National University of Singapore, Singapore 117543, Singapore.
Samira Husen
SH
Alamudi
Department of Chemistry &Med Chem Program, National University of Singapore, Singapore 117543, Singapore.
Rudrakanta
R
Satapathy
Department of Chemistry, Korea Advanced Institute of Science and Technology, Daejeon 305701, Republic of Korea.
Jihyo
J
Kim
Laboratory of Bioimaging Probe Development, Singapore Bioimaging Consortium, Agency for Science, Technology and Research, Singapore 138667, Singapore.
Dongdong
D
Su
Department of Pharmaceutical Chemistry and the Cardiovascular Research Institute, University of California San Francisco, California 94158, USA.
Haiyan
H
Ren
Department of Chemistry &Med Chem Program, National University of Singapore, Singapore 117543, Singapore.
Rajkumar
R
Das
Department of Chemistry &Med Chem Program, National University of Singapore, Singapore 117543, Singapore.
Lingna
L
Hu
Departamento de Química, Universidad de Guanajuato, Guanajuato 36050, Mexico.
Enrique
E
Alvarado-Martínez
Department of Chemistry &Med Chem Program, National University of Singapore, Singapore 117543, Singapore.
Jung Yeol
JY
Lee
Department of Pharmaceutical Chemistry and the Cardiovascular Research Institute, University of California San Francisco, California 94158, USA.
Christian
C
Hoppmann
Departamento de Química, Universidad de Guanajuato, Guanajuato 36050, Mexico.
Eduardo
E
Peña-Cabrera
College of Pharmacy and Research Institute of Life and Pharmaceutical Sciences, Sunchon National University, Sunchon 540950, Republic of Korea.
Hyung-Ho
HH
Ha
Department of Chemistry, Korea Advanced Institute of Science and Technology, Daejeon 305701, Republic of Korea.
Hee-Sung
HS
Park
Department of Pharmaceutical Chemistry and the Cardiovascular Research Institute, University of California San Francisco, California 94158, USA.
Lei
0000-0002-5859-2526
L
Wang
Department of Chemistry &Med Chem Program, National University of Singapore, Singapore 117543, Singapore.|Laboratory of Bioimaging Probe Development, Singapore Bioimaging Consortium, Agency for Science, Technology and Research, Singapore 138667, Singapore.
Young-Tae
YT
Chang
D000818:Animals; D001386:Azides / Q000737:chemistry*; D001896:Boron Compounds / Q000737:chemistry*; D016466:CHO Cells; D045744:Cell Line, Tumor; D002466:Cell Nucleolus / Q000378:metabolism / Q000648:ultrastructure; D003412:Cricetulus; D034242:Cyclooctanes / Q000737:chemistry*; D015195:Drug Design; D005456:Fluorescent Dyes / Q000138:chemical synthesis* / Q000378:metabolism; D015870:Gene Expression; D006056:Golgi Apparatus / Q000378:metabolism / Q000648:ultrastructure; D057166:High-Throughput Screening Assays; D006801:Humans; D008247:Lysosomes / Q000378:metabolism / Q000648:ultrastructure; D008928:Mitochondria / Q000378:metabolism / Q000648:ultrastructure; D057054:Molecular Imaging / Q000379:methods*; D010006:Osteoblasts / Q000378:metabolism / Q000648:ultrastructure; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D059629:Signal-To-Noise Ratio; D013194:Staining and Labeling / Q000379:methods*
C095489:4,4-difluoro-4-bora-3a,4a-diaza-s-indacene; D001386:Azides; D001896:Boron Compounds; D034242:Cyclooctanes; D005456:Fluorescent Dyes; D011994:Recombinant Proteins