Liu Wenqian, Pan Yingjia, Zhang Yu, Dong Chang, Huang Lei, Lian Jiazhang
Key Laboratory of Biomass Chemical Engineering of Ministry of Education and State Key Laboratory of Biobased Transportation Fuel Technology, College of Chemical and Biological Engineering, Zhejiang University, Hangzhou 310027, China; BGI Research, Hangzhou 310030, China.
Key Laboratory of Biomass Chemical Engineering of Ministry of Education and State Key Laboratory of Biobased Transportation Fuel Technology, College of Chemical and Biological Engineering, Zhejiang University, Hangzhou 310027, China; ZJU-Hangzhou Global Scientific and Technological Innovation Center, Zhejiang University, Hangzhou 310000, China.
Trends Biotechnol. 2025 Jun;43(6):1356-1370. doi: 10.1016/j.tibtech.2024.10.011. Epub 2024 Nov 12.
Despite the prevalence of genome editing tools, there are still some limitations in dynamic and continuous genome editing. In vivo single-stranded DNA (ssDNA)-mediated genome mutation has emerged as a valuable and promising approach for continuous genome editing. In this review, we summarize the various types of intracellular ssDNA production systems and notable achievements in genome engineering in both prokaryotic and eukaryotic cells. We also review progress in the development of applications based on retron-based systems, which have demonstrated significant potential in molecular recording, multiplex genome editing, high-throughput functional variant screening, and gene-specific continuous in vivo evolution. Furthermore, we discuss the major challenges of ssDNA-mediated continuous genome editing and its prospects for future applications.
尽管基因组编辑工具已广泛应用，但在动态和连续基因组编辑方面仍存在一些局限性。体内单链DNA（ssDNA）介导的基因组突变已成为一种有价值且前景广阔的连续基因组编辑方法。在本综述中，我们总结了细胞内ssDNA产生系统的各种类型以及原核和真核细胞基因组工程中的显著成就。我们还回顾了基于反转录子系统的应用开发进展，该系统在分子记录、多重基因组编辑、高通量功能变体筛选以及基因特异性体内连续进化方面已展现出巨大潜力。此外，我们讨论了ssDNA介导的连续基因组编辑的主要挑战及其未来应用前景。
