Human apurinic/apyrimidinic endonuclease/redox effector factor 1 (APE1) is an essential DNA repair protein. Herein, we demonstrate that avidin-oriented abasic site-containing DNA strands (AP-DNA) on the surface of silica coated magnetic nanoparticles (SiMNP) can selectively respond to APE1 while resist the digestion by other nucleases. Mechanism studies have revealed that avidin may serve as an organizer protein and recruit APE1 to the DNA substrates on the nanoparticles via strong and specific interactions. Taking advantage of this newly disclosed property, we for the first time successfully displayed the intracellular activities of APE1 in living cells by fluorescence imaging. The avidin organized AP-DNA-SiMNP assembly holds great potential for enzyme-mediated release of drugs inside tumor cells which often contain higher levels of APE1 than normal cells.

We demonstrate the successful construction of fluorescently labeled magnetic antibody-like nanoparticles (ANPs) via a facile one-step surface-initiated in situ molecular imprinting approach over silica coated magnetite (Fe3O4@SiO2) core-shell nanocomposites. The as-prepared ANPs had a highly compact structure with an overall size of 83 ± 5 nm in diameter and showed excellent aqueous dispersion stability. With the predetermined high specificity to the target protein and high biocompatibility, the ANPs enabled rapid, efficient, selective and optically trackable sequestration of target proteins within living cells. This work represents the first example of fully artificially engineered multifunctional ANPs for the intracellular protein-sequestration without disruption of the cells. The established approach may be further extended to generate ANPs for various proteins of interest and provide useful tools for related biological research and biomedical applications.

A novel DNA structure containing a 3' internal-loop modified abasic site has been constructed which enables effective differentiation between apurinic/apyrimidinic endonuclease (APE1) and nonspecific endonuclease (DNase I). When this unique substrate structure is employed, a double-loop frayed-end chimeric fluorescent probe is successfully developed for quantitative measurement of the activity of APE1 in biological samples without the need of additional cleanup or preconcentration steps. The method is simple and rapid and has a single-step with a linear working range between 0.1 and 5.0 U/mL and a lower limit of detection of 0.1 U/mL. It holds great potential in real-time monitoring of the variation of intracellular and extracellular APE1, which will be very useful for further understanding of the DNA repair pathways in different organisms.