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. 2010 Jan 4;11(1):47-53.
doi: 10.1002/cbic.200900529.

Principles and applications of the photochemical control of cellular processes

Alexander Deiters  1
Affiliations

Principles and applications of the photochemical control of cellular processes

Alexander Deiters. Chembiochem. .
No abstract available

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Figures

Figure 1
Figure 1
Light-activated phosphorothioate antisense agents. A) Caged thymidine nucleotide incorporated into an antisense PS DNA. The caging group (blue) is removed through UV irradiation; this enables hybridization of the antisense agent to the mRNA, and thus leads to gene silencing. B) Spatial regulation of Renilla luciferase expression by using caged PS DNA antisense agents. The cellular monolayer was irradiated only inside the white circle (365 nm, 5 min, 23 W); this led to localized silencing of luciferase in cells transfected with the caged PS DNA, compared to C) cells which have not been irradiated. Adapted with permission from: ChemBioChem 2008, 9, 2937–2940. Copyright Wiley-VCH Verlag GmbH & Co. KGaA.
Figure 2
Figure 2
A) Exposure of a single-skinned cardiac trabecula to the caged calcium 1 (1 mm), followed by flash photolysis. B)–C) Identical experiments performed by using a classical ortho-nitrobenzyl analogue of 1 and one or two irradiations; this led to lower tension recordings. Adapted by permission from Macmillan Publishers Ltd: Nat. Methods 2006, 3, 35–40, Copyright 2006.
Figure 3
Figure 3
Cre recombinase was rendered completely inactive through site-specific incorporation of an ortho-nitrobenzyl- protected tyrosine residue at position Y324. Irradiation with UV light (365 nm, 5 min) restores activity and triggers DNA recombination in HEK293T cells. Photochemical activation of GFP and simultaneous deactivation of DsRed was achieved in mammalian cell culture through the excision of the DsRed gene and a stop codon through DNA recombination between two loxP sites (black triangles). CMV=cytomegalovirus promoter.
Figure 4
Figure 4
Potential scenarios for the biological activity of caged molecules before (■) and after (●) irradiation. The biological activity increases from blue to red; however, a cellular environment often displays a threshold of biological activity to be reached before activation of the desired biological effect.
Figure 5
Figure 5
A) Light-irradiation of the caged IPTG (2) followed by intracellular hydrolysis of ester 3 to yield IPTG (4). B) Bacterial lithography with UV irradiation of 365 nm for 30 s while blocking the left half of a Petri dish. Two different reporter genes were employed; lacZ (left) and GFP (right) display a negative and a positive image, respectively. Adapted with permission from: Angew. Chem. Int. Ed. 2007, 46, 4290–4292. Copyright Wiley-VCH Verlag GmbH & Co. KGaA.
Figure 6
Figure 6
A) Schematic representation of the reversible LOV–Rac1 light-regulation. B) Pull-down assays of selected LOV–Rac1 mutants in the dark. C) Irradiation of HeLa cells expressing LOV–Rac1 induced lamellipodial protrusions and membrane ruffles within minutes. D) Spatially restricted irradiation enables localized control of Pac1 activity. Adapted by permission from Macmillan Publishers Ltd: Nature 2009, 461, 104–108, copyright 2009.
Figure 7
Figure 7
Photoactivated gene function in a mouse embryo by using caged doxycycline as a small molecule inducer of gene expression. A) Fluorescence image of widespread GFP fluorescence in a control embryo at day 10.5, incubated with 20 µm doxycycline. B) Fluorescence image of an embryo incubated with 2.6 µm of caged doxycycline without irradiation, C) after a single 15 s pulse of UV irradiation, and D) after a second irradiation pulse 3 h later. Adapted by permission from Macmillan Publishers Ltd: Nat. Methods 2009, 6, 527–531, copyright 2009.
Scheme 1
Scheme 1
General decaging reaction of a molecule (sphere), caged with a classical ortho-nitrobenzyl group. Chemical and photochemical properties of the caging group can be tuned with suitable substituents, including: R1=H, OCH3; R2=H, CH3, CO2H. X=O, CO2, S, NH, NHCO2, OPO3H.
See this image and copyright information in PMC

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