Li Jieyi, Tong Hanze, Wu Yi
School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, China.
Frontier Science Center for Synthetic Biology and Key Laboratory of Systems Bioengineering, Ministry of Education, Tianjin University, Tianjin 300072, China.
Sheng Wu Gong Cheng Xue Bao. 2023 Jun 25;39(6):2465-2484. doi: 10.13345/j.cjb.230278.
Large-scale genetic manipulation of the genome refers to the genetic modification of large fragments of DNA using knockout, integration and translocation. Compared to small-scale gene editing, large-scale genetic manipulation of the genome allows for the simultaneous modification of more genetic information, which is important for understanding the complex mechanisms such as multigene interactions. At the same time, large-scale genetic manipulation of the genome allows for larger-scale design and reconstruction of the genome, and even the creation of entirely new genomes, with great potential in reconstructing complex functions. Yeast is an important eukaryotic model organism that is widely used because of its safety and easiness of manipulation. This paper systematically summarizes the toolkit for large-scale genetic manipulation of the yeast genome, including recombinase-mediated large-scale manipulation, nuclease-mediated large-scale manipulation, synthesis of large DNA fragments and other large-scale manipulation tools, and introduces their basic working principles and typical application cases. Finally, the challenges and developments in large-scale genetic manipulation are presented.
基因组的大规模遗传操作是指利用基因敲除、整合和易位对大片段DNA进行基因改造。与小规模基因编辑相比,基因组的大规模遗传操作能够同时修改更多的遗传信息,这对于理解多基因相互作用等复杂机制至关重要。同时,基因组的大规模遗传操作能够对基因组进行更大规模的设计和重建,甚至创造全新的基因组,在重建复杂功能方面具有巨大潜力。酵母是一种重要的真核模式生物,因其安全性高且易于操作而被广泛应用。本文系统总结了酵母基因组大规模遗传操作的工具包,包括重组酶介导的大规模操作、核酸酶介导的大规模操作、大DNA片段合成及其他大规模操作工具,并介绍了它们的基本工作原理和典型应用案例。最后,阐述了大规模遗传操作面临的挑战和发展趋势。