DNA has been demonstrated to be a promising engineering material for the fabrication of pre‐designed nanostructures and nanodevices, which feature structural programmability, spatial addressability, dynamic mechanics, and biocompatibility. Here, we review the recent progress in the design, construction and biomedical applications of DNA nanostructures, particularly imaging probes and drug delivery carriers. We discuss several key points for in vivo biomedical applications of DNA‐based delivery systems, such as the cost and safety issues. In the end, we explore the challenges and opportunities of DNA nanotechnology in the field of nanomedicine.