Abstract

As critical regulators of gene expression, microRNAs (miRNAs) are key biomarkers and therapeutic targets in cancer. However, current methods for intracellular miRNA imaging are often limited by poor sensitivity and operational complexity. In this study, we identified a site-specifically modified DNAzyme variant, 11Bn, which exhibits up to 7-fold higher catalytic activity than the wild-type 8-17 through systematic screening. Using this variant, we constructed a DNAzyme-based sensor for miRNA-21 imaging in living cells. The sensor achieves a limit of detection of 7.89 nM, threefold lower than that of the wild-type sensor, and enables sensitive visualization of intracellular miRNA-21 without signal amplification. Moreover, it can capture dynamic changes in miRNA levels within cells, providing a versatile molecular tool for miRNA imaging and related biomedical applications.