Results: 4

1.
Figure 2

Figure 2. From: Temporal and spatial regulation of microRNA activity with photo-activatable cantimirs.

In vitro characterization of lsy-6 cantimirs. (A) Structures and mass analyses of lsy-6 cantimirs containing blocking strands of different lengths. (B) UV-dose dependent photolysis of canti_lsy-6_11 analyzed by the gel shift assay. (C) Time courses of photo-conversion of canti_lsy-6_11 and a reference caged coumarin, NPE-HCC (ref. 19).

Genhua Zheng, et al. ACS Chem Biol. 2011 December 16;6(12):1332-1338.
2.
Figure 1

Figure 1. From: Temporal and spatial regulation of microRNA activity with photo-activatable cantimirs.

Design and synthesis of caged antimir (cantimir). (A) Design of cantimir for the light controlled inhibition of microRNA. The black and blue square represents caged coumarin (nonfluorescent) and coumarin, respectively. (B) Structure of a bifunctional caged linker. (C) Synthesis of cantimirs against lsy-6 miRNA. (a) 5 mM tris(2-carboxyethyl)phosphine, phosphate buffer(50 mM, pH 7.0), argon, 1 hr, 95%. (b) 0.1 M caged linker in DMSO, 3 mM oligoribonucleotide in phosphate buffer (100 mM, pH 7.24), 2: 1 (v/v); 35 °C, 35 minutes, 85%. (c) Water, room temperature, overnight, 31%.

Genhua Zheng, et al. ACS Chem Biol. 2011 December 16;6(12):1332-1338.
3.
Figure 4

Figure 4. From: Temporal and spatial regulation of microRNA activity with photo-activatable cantimirs.

Stage and cell specific inhibition of lsy-6 in developing C. elegans. (A) Worms (gcy-5::gfp) labeled with canti_lsy-6_11 were photolyzed with UV light (365 nm) at five developmental stages. The lsy-6(lf) phenotype was scored similarly as in Figure 3. (B) Cell specific inhibition of lsy-6 in developing C. elegans. Local photolysis of canti_lsy-6_11 in ABa (a, b), but not in ABp (c, d), of 4-cell stage embryos resulted in lsy-6(lf) in the adult worms. Fluorescence images (b, d) of 4-cell embryos during local photolysis where a restricted beam of UV light (indicated by the yellow circle) was shone only on the selected cell. N = 12 for each condition.

Genhua Zheng, et al. ACS Chem Biol. 2011 December 16;6(12):1332-1338.
4.
Figure 3

Figure 3. From: Temporal and spatial regulation of microRNA activity with photo-activatable cantimirs.

In vivo assay of the background and light-activated antisense activity of lsy-6 cantimirs. (A) lsy-6 mediates inhibition of gcy-5 gene expression in the ASE class of chemosensory neurons19. A simplified version of a more complex gene regulatory network23 is shown. (B) Example fluorescence images of gcy-5prom::gfp expression in ASE neurons. (C) Embryos labeled with lsy-6 cantimirs with different blocking strands were either kept in the dark (no UV) or illuminated with 365 nm UV light (UV) prior to the comma stage and scored for the lsy-6(lf) phenotype when they reached adults. All the experiments were repeated at least twice, and in each time at least 50 worms were scored. The error bars are standard deviations.

Genhua Zheng, et al. ACS Chem Biol. 2011 December 16;6(12):1332-1338.

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