Skip to main content
. 2022 Oct 18;144(43):19791–19798. doi: 10.1021/jacs.2c06599

Figure 3.

Figure 3

Delayed SDR using Fpg and UDG enzymes. (a) Scheme showing temporal control of SDR using the enzyme Fpg and a blocker strand containing a modified Goxo base in its center. (b) Time-course experiments in the presence of the Goxo-blocker (150 nM) after the addition of different concentrations of Fpg. (c) Half-life values vs Fpg concentrations at a fixed concentration of the Goxo-blocker strand (150 nM). (d) Half-life values vs Goxo-blocker concentrations at a fixed concentration of Fpg (8.0 U/mL). (e) Scheme showing temporal control of SDR using the enzyme UDG and a blocker strand containing uracil bases. (f) Time-course experiments in the presence of the uracil-blocker (150 nM) after the addition of different concentrations of UDG. (g) Half-life values vs UDG concentrations at a fixed concentration of the uracil-blocker strand (150 nM). (h) Half-life values vs uracil blocker concentrations at a fixed concentration of UDG (0.05 U/mL). In (b–d) and (f–h), experimental values (dots) are shown together with fits (b,f) and prediction (c,d,g,h) from a parameterized kinetic model (solid lines) described in section 2e,f of Supporting Information. Experiments were performed in Tris HCl 20 mM, MgCl2 10 mM, EDTA 1 mM, pH 8.0 at T = 30 °C. [Target duplex] = 50 nM, [input strand] = 50 nM.