An aptamer-based ligation-triggered rolling circle amplification strategy for ATP detection and imaging in situ

Abstract Adenosine 5′-triphosphate (ATP), one of the most widely known small natural biological molecules, is a multifunctional molecule existing in living organisms and plays an important role in many biological processes. In this work, we have developed an aptamer-based ligation-triggered rolling circle amplification strategy for ATP detection. Based on ATP aptamer, phosphorylated single oligonucleotide could be specifically ligated and circularized in the presence of ATP. Then branched rolling circle amplification (BRCA) was used as a signal amplification method for the analysis of ATP. Due to the high binding affinity and specificity of ATP aptamer, the proposed method showed significant advantages in high sensitivity and excellent selectivity. In addition, the selectivity was further improved in the ligation step by T4 ligase, it can also distinguish ATP from other adenosine analogues. Furthermore, this simple and facile method could be used to detect ATP in complex biological matrix, cell lysates, and the detection limit was as low as five cells. Moreover, the strategy realized the visualization of ATP level in situ and providing the location of ATP at single-cell level.