Click-Type Protein-DNA Conjugation for Mn2+ Imaging in Living Cells

Abstract

A click-type protein-DNA conjugation, named as MnDDC (Mn²⁺-activated DCV-DNA conjunction), is presented, where DCV (rep protein of duck circovirus) and its target DNA work as the modular blocks to rapidly and effectively generate Mn²⁺-dependent and site-specific protein-DNA linkage. On the basis of MnDCC, a fluorescent Mn²⁺ biosensor composed of DCV and a molecular beacon, was developed for rapid sensing of Mn²⁺ within 2 min with nanomolar sensitivity. Using the proposed biosensor, not only analysis of Mn²⁺ in real samples (e.g., serum and food), but also wash-free fluorescent imaging of Mn²⁺ in extracellular environment and cytoplasm have been achieved. Moreover, the MnDDC-based sensor was proved to be a powerful tool for visualization of Mn²⁺ during exploration of the associated cytotoxicity in living neural cells, which is helpful to reveal the cellular responses toward the disordered homeostasis of Mn²⁺ in both extracellular and intracellular microenvironments.