Wang Xiaotong, Xu Guangxue, Johnson William A, Qu Yuanhao, Yin Di, Ramkissoon Nurupa, Xiang Hong, Cong Le
Department of Pathology, Stanford University School of Medicine, Stanford, CA 94305, USA.
Department of Genetics, Stanford University School of Medicine, Stanford, CA 94305, USA.
Curr Opin Biomed Eng. 2023 Dec;28. doi: 10.1016/j.cobme.2023.100491. Epub 2023 Jul 22.
CRISPR/Cas-based gene-editing technologies have emerged as one of the most transformative tools in genome science over the past decade, providing unprecedented possibilities for both fundamental and translational research. Following the initial wave of innovations for gene knock-out, epigenetic/RNA modulation, and nickase-mediated base-editing, recent efforts have pivoted towards long-sequence gene editing- specifically, the insertion of large fragments (>1 kb) into the endogenous genome. In this review, we survey the development of these CRISPR/Cas-based sequence insertion methodologies in conjunction with the emergence of novel families of editing enzymes, such as transposases, single-stranded DNA-annealing proteins, recombinases, and integrases. Despite facing a number of challenges, this field continues to evolve rapidly and holds the potential to catalyze a new wave of revolutionary biomedical applications.
在过去十年中，基于CRISPR/Cas的基因编辑技术已成为基因组科学中最具变革性的工具之一，为基础研究和转化研究提供了前所未有的可能性。继基因敲除、表观遗传/RNA调控和切口酶介导的碱基编辑的首轮创新之后，最近的研究工作已转向长序列基因编辑——具体而言，是将大片段（>1 kb）插入内源性基因组。在本综述中，我们结合转座酶、单链DNA退火蛋白、重组酶和整合酶等新型编辑酶家族的出现，综述了这些基于CRISPR/Cas的序列插入方法的发展。尽管面临诸多挑战，但该领域仍在迅速发展，并有潜力催生出新一轮革命性的生物医学应用。
