Fig. 3: Detection of modifications on nanolatches.
a, A schematic depicting how 5mC modification influences C/G base pairing before and after bisulfite conversion. The MS2 carrier design is identical to that in Fig. 2. b, The nanopore measurement results of MS2 carriers incubated with nanolatches containing 0–4 5mC modifications, with (+) and without (−) bisulfite treatment. c, A schematic of how inosine modification affects A/U base pairing, along with nanopore measurement results for nanolatches containing 0–4 inosines in the sequence. d, A schematic of MeC modification and nanopore measurement results for nanolatches possessing 0–4 MeCs. The data in b–d are shown as the mean ± standard deviation from three independent measurements. e, A theoretical model of the nanolatch system for detecting modifications. The presence of modifications can either strengthen or weaken the stability of the duplex between the nanolatch and the RNA scaffold, altering the energy difference \(\Delta G\) between ‘latched’ and ‘unlatched’ energy levels. R is the ideal gas constant, T is the absolute temperature at which the reaction occurs, \({K}_{n}\) is the equilibrium constant of loop latching, and \(n\) is the number of modifications. f, The linear fitting results of \(\mathrm{ln}({K}_{n})\) against \(n\) for each type of modifications studied in this work. 5mC (+) and 5mC (−) refer to cytosine methylations with (+) and without (−) bisulfite treatment, respectively. The data point for four inosines (open square) was excluded from fitting, as the positive event ratio had already dropped to nearly 0% with three inosines.
