Simultaneous ultrasensitive ADP and ATP quantification based on CRISPR/Cas12a integrated ZIF-90@Ag₃AuS₂@Fe₃O₄...

ADP/ATP ratio is a sensitive indicator of changes in cellular energy status and is important for regulating cell signaling activities. Ultrasensitive quantification of ADP and ATP concentration in a single system is in great demand for bioanalysis and early disease diagnosis. Hence, a target-regulated luminous nanoplatform based on clustered regularly interspaced short palindromic repeats (CRISPR)/Cas12a integrated zeolite imidazolate framework (ZIF-90)@Ag₃AuS₂@Fe₃O₄ nanocomposites was established for the simultaneous detection of ADP and ATP. This simultaneous and ultrasensitive quantification nanoplatform (dsDNA-ZIF-90@Ag₃AuS₂@Fe₃O₄) composed an ADP sensitive module based on the aptamer-activated CRISPR/Cas12a and an ATP responsive module based on ATP-triggered ZIF-90 decomposition and quencher loading release. The binding and signaling processes of the different nucleotides were independent, and there was no interference between the two modules. Utilizing the high specific binding and strong signal amplification of this method, limits of detection as low as 0.022 and 0.079 nM were obtained within 30 min for ADP and ATP, respectively. Moreover, the proposed biosensor exhibited high accuracy, specificity, and excellent recovery in serum samples and bacterial biofilms. Therefore, the dsDNA-ZIF-90@Ag₃AuS₂@Fe₃O₄-based nanoplatform provides a promising method for ultrasensitive dual-mode quantification of ADP and ATP in the same system, possessing great potential for bioanalysis and early disease diagnosis.