Fig. 6: Demonstration of the CRISPR MN wearable system in mice.
a Timeline of real-time monitoring in CNE-Luc-bearing mice. b Schematic illustration of in vivo real-time sampling and monitoring, including (I) chemiluminescence bioimaging and (II) CRISPR MN for CNE-Luc-bearing mice. The CRISPR MN system was calibrated in PBS (37 °C, 0.01 M, pH 7.4) for 3 min to eliminate sensor-to-sensor variation in electrical output. The red circle represented the detection time point. The skin stratum corneum of all BALB/c nude mice was cleaned by scrub cream, disinfection with 75% ethanol, and smearing with 100 μL TE buffer (pH 8.0). The region of interest on mouse skin was dried with cotton. Finally, the mice were placed on a heat plate during the real-time monitoring procedures. c Parallel trials on mice at different time points, including 2, 8, 24, 48, 72, and 120 h, scale bar: 1 cm. d Slope values collected from the curve peak of BALB/c nude mice real-time curve calculated by simple differentiation, including 2, 8, 72, 120 h, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, as analyzed by two-way ANOVA, p value of 0.00011, 0.0000076, 0.000042, and 0.0045 for 2, 8, 72, and 120 h, respectively, data presented as mean values ± SD, n = 3 biologically independent animals. e In vivo dynamic change in EBV cfDNA levels in BALB/c nude mice detected by the CRISPR MN system during the first 5 days, the curve was fitted by spline curve mode, Origin 2018 software. f Four independent methods applied to 18-day CNE-Luc-bearing BALB/c nude mice demonstrated that the CRISPR MN wearable system was as accurate as the gold-standard PCR (blood sampling by a commercial kit) in terms of qualitative analysis. g The stable sensitivity of the CRISPR MN in vivo, the CRISPR MN laminated on BALB/c nude mice, then transferred to a skin chip for monitoring 3 × 10−14 M target cfDNA under reverse iontophoresis (10 V), data presented as mean values ± SD, n = 3 independent experiments.
