DNA cleavage plays a crucial role in many biological processes such as DNA replication, transcription, and recombination. It is also a powerful tool in gene editing, therapeutics and biosensor design. Nanozymes aim to develop nanomaterial-based enzyme mimics. Compared with natural enzymes, nanozymes offer advantages of higher stability, lower cost, and recyclability. Recently, nanozymes with interesting DNA cleavage activities have emerged, including both hydrolytic and oxidative cleavage. This Perspective starts by introducing DNA cleavage of nanozymes, focusing on recent examples. Some interesting nanozymes include CeO2 nanoparticles for the hydrolytic cleavage of single-stranded DNA oligonucleotides, chiral carbon dots mimicking topoisomerase activity, and light-assisted cleavage of DNA. The corresponding cleavage mechanisms are then discussed along with a few representative applications for DNA repair and as antibacterial agents. Finally, a few future research opportunities are discussed.