Park Sun-Ji, Lee Ga Eun, Cho Soo Min, Choi Eui-Hwan
New Drug Development Center, Daegu-Gyeongbuk Medical Innovation Foundation, Daegu 41061, South Korea.
Department of Biotechnology, Korea National University of Transportation, Chungbuk 27909, South Korea.
Mol Ther Nucleic Acids. 2025 Jul 17;36(3):102634. doi: 10.1016/j.omtn.2025.102634. eCollection 2025 Sep 9.
The CRISPR-Cas system has transformed our ability to edit and modify genomes in eukaryotic cells, offering unmatched precision and broad applicability. By utilizing a programmable RNA protein complex to introduce targeted double-strand breaks, the CRISPR-Cas system enables the correction of pathogenic mutations and the modulation of gene function with unprecedented efficiency. Its broad applicability spans the correction of inherited genetic defects through homology-directed repair to the disruption of deleterious alleles via non-homologous end joining. In this review, we first outline the molecular architecture and mechanistic basis of CRISPR-Cas9 and then consider its latest applications in modeling, drug screening, small-molecule-mediated editing, and treating hereditary, autoimmune, and oncological diseases. Emphasis is placed on the generation of disease-relevant cellular and animal models and on the potential of CRISPR-Cas9-mediated gene therapy to address hitherto intractable disorders. Finally, we discuss current challenges including off-target activity, gene editing efficiency, delivery constraints, and immunogenicity and highlight emerging strategies to overcome these hurdles and broaden the clinical impact of CRISPR-Cas systems.
CRISPR-Cas系统改变了我们编辑和修饰真核细胞基因组的能力，具有无与伦比的精准度和广泛的适用性。通过利用一种可编程的RNA-蛋白质复合物引入靶向双链断裂，CRISPR-Cas系统能够以前所未有的效率纠正致病突变并调节基因功能。其广泛的适用性涵盖了从通过同源定向修复纠正遗传性基因缺陷到通过非同源末端连接破坏有害等位基因。在本综述中，我们首先概述CRISPR-Cas9的分子结构和作用机制，然后探讨其在疾病模型构建、药物筛选、小分子介导的编辑以及治疗遗传性、自身免疫性和肿瘤性疾病方面的最新应用。重点在于生成与疾病相关的细胞和动物模型以及CRISPR-Cas9介导的基因疗法解决迄今难以治疗的疾病的潜力。最后，我们讨论当前面临的挑战，包括脱靶活性、基因编辑效率、递送限制和免疫原性，并强调克服这些障碍和扩大CRISPR-Cas系统临床影响的新兴策略。
