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. 2023 Jun 2;24(13):e202200756. doi: 10.1002/cbic.202200756

Figure 4.

Figure 4

A) Late‐stage modification of oligonucleotides. Alkyne‐containing compounds 12 a–13 a, 15 a–22 a and 24 a were reacted with 5’‐azido oligonucleotide 25 in a CuAAC reaction to give modified oligonucleotides 12 b–13 b, 15 b–22 b and 24 b. B) Evaluation of recognition of modified oligonucleotides 12 b–13 b, 15 b–22 b and 24 b by SNM1A. Digestion of fluorescent substrate 11 (0.8 pmol) after incubation with SNM1A (50 fmol) for 60 min at 37 °C following 5 min of preincubation with oligonucleotides 12 b–13 b, 15 b–22 b and 24 b (2.8 μM). Duplicate data are available in Figure S2. nt=nucleotides; sequence of 5’‐azido oligonucleotide 25: 5′‐(azideT)AG CAG TCA GTC AGT CAT GC‐3′; sequence of fluorescent substrate oligonucleotide 11: 5’‐XTAG CAG TCA GTC AGT CAT CGY‐3’, X=thymidine 5’‐phosphate, Y=Cy3.