Kolesova Ekaterina, Pulone Sabina, Kostyushev Dmitry, Tasciotti Ennio
Scientific Center for Translation Medicine, Sirius University of Science and Technology, Sochi 354340, Russia.
Human Longevity Program, IRCCS San Raffaele Roma, Rome, Italy.
Adv Drug Deliv Rev. 2025 Sep;224:115619. doi: 10.1016/j.addr.2025.115619. Epub 2025 May 29.
This review explores the transformative role of CRISPR/Cas systems in optical bioimaging, emphasizing how advancements in nanoparticle (NP) technologies are revolutionizing the visualization of gene-editing processes both in vitro and in vivo. Optical imaging techniques, such as near-infrared (NIR) and fluorescence imaging, have greatly benefited from the integration of nanoformulated contrast agents, improving resolution, sensitivity, and specificity. CRISPR/Cas systems, originally developed just for gene editing, are now being coupled with these imaging modalities to enable real-time monitoring and quantitative measurements of metabolites, vitamins, proteins, nucleic acids and other entities in specific areas of the body, as well as tracking of CRISPR/Cas delivery, editing efficiency, and potential off-target effects. The development of CRISPR/Cas-loaded NPs allows for enhanced imaging and precise monitoring across multiple scales with multiplexed and multicolor imaging in complex settings, including potential in vivo diagnostics. CRISPR/Cas therapeutics as well as diagnostics are hindered by the lack of efficient and targeted delivery tools. Biomimetic NPs have emerged as promising tools for improving biocompatibility, enhancing targeting capabilities, and overcoming biological barriers, facilitating more efficient delivery and bioimaging of CRISPR/Cas systems in vivo. As the design of these NPs and delivery mechanisms improves, alongside advancements in endolysosomal escape, CRISPR/Cas-based bioimaging will continue to advance, offering unprecedented possibilities in precision medicine and theranostic applications.
本综述探讨了CRISPR/Cas系统在光学生物成像中的变革性作用，强调了纳米颗粒（NP）技术的进步如何正在彻底改变体外和体内基因编辑过程的可视化。光学成像技术，如近红外（NIR）和荧光成像，因纳米配方造影剂的整合而受益匪浅，提高了分辨率、灵敏度和特异性。CRISPR/Cas系统最初仅用于基因编辑，现在正与这些成像方式相结合，以实现对体内特定区域的代谢物、维生素、蛋白质、核酸和其他实体的实时监测和定量测量，以及对CRISPR/Cas递送、编辑效率和潜在脱靶效应的追踪。负载CRISPR/Cas的NP的开发使得在复杂环境中通过多重和多色成像在多个尺度上实现增强成像和精确监测成为可能，包括潜在的体内诊断。缺乏高效且有针对性的递送工具阻碍了CRISPR/Cas疗法以及诊断方法的发展。仿生NP已成为有前景的工具，可用于改善生物相容性、增强靶向能力和克服生物屏障，促进CRISPR/Cas系统在体内更高效的递送和生物成像。随着这些NP的设计和递送机制的改进，以及内溶酶体逃逸方面的进展，基于CRISPR/Cas的生物成像将继续发展，在精准医学和治疗诊断应用中提供前所未有的可能性。
