Development of background-free tame fluorescent probes for intracellular live cell imaging View Full Text


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

2016-06-20

AUTHORS

Samira Husen Alamudi, Rudrakanta Satapathy, Jihyo Kim, Dongdong Su, Haiyan Ren, Rajkumar Das, Lingna Hu, Enrique Alvarado-Martínez, Jung Yeol Lee, Christian Hoppmann, Eduardo Peña-Cabrera, Hyung-Ho Ha, Hee-Sung Park, Lei Wang, Young-Tae Chang

ABSTRACT

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. More... »

PAGES

11964

Identifiers

URI

http://scigraph.springernature.com/pub.10.1038/ncomms11964

DOI

http://dx.doi.org/10.1038/ncomms11964

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1007118825

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/27321135


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30 schema:description 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.
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37 schema:keywords AzG-1
38 BODIPY probe
39 Co 1
40 Waals surface area
41 area
42 azide-containing BODIPY probe
43 background
44 background-free probe
45 background-free probe development
46 background-free tame fluorescent probes
47 biomolecule's roles
48 biomolecules
49 cell imaging
50 cell intracellular imaging
51 cell permeable background-free probe development
52 cells
53 cellular components
54 cheminformatics
55 combination
56 components
57 cyclooctyne
58 depth understanding
59 der Waals surface area
60 design
61 desirable labelling probes
62 development
63 efficient strategy
64 engineered proteins
65 first predictive model
66 fluorescence labeling
67 fluorescent probe
68 function
69 high signal
70 high-throughput screening
71 imaging
72 interaction
73 intracellular biomolecules
74 intracellular imaging
75 intracellular live cell imaging
76 intracellular target organelles
77 labeling
78 labeling probes
79 lipophilicity
80 live cell intracellular imaging
81 live-cell imaging
82 living cells
83 minimum background
84 model
85 native living cells
86 noise ratio
87 nonspecific interactions
88 novel tool
89 organelles
90 permeable background-free probe development
91 powerful strategy
92 predictive model
93 probe
94 probe development
95 protein
96 ratio
97 rational design
98 reliability
99 results
100 role
101 screening
102 signals
103 solubility
104 strategies
105 such probes
106 surface area
107 tame fluorescent probes
108 target organelles
109 tool
110 understanding
111 van der Waals surface area
112 water solubility
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