酵母基因组大尺度遗传操纵工具研究进展  

Tools for large-scale genetic manipulation of yeast genome

作　　者：李杰奕 佟函泽 吴毅 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)

机构地区：[1]天津大学化工学院,天津300072 [2]天津大学教育部合成生物学前沿科学中心和系统生物工程重点实验室,天津300072

出　　处：《生物工程学报》2023年第6期2465-2484,共20页Chinese Journal of Biotechnology

基　　金：国家自然科学基金(31971351);国家重点研发计划(2021YFC2102500)。

摘　　要：基因组大尺度遗传操纵是指对基因组大片段DNA的敲除、整合、易位等遗传改造。相较于小规模基因编辑,基因组大尺度遗传操纵可实现更多遗传信息的同步改造,对于探究多基因相互作用等复杂机制的理解有重要意义。同时,基因组大尺度遗传操纵技术可对基因组开展更大规模的设计重构,甚至创建全新的基因组,在复杂功能重塑方面具有重要创新潜力。酵母是一种重要的真核模式生物,因其安全性和易于操作而被广泛应用。本文系统总结了酵母基因组大尺度遗传操纵的工具包,包括重组酶介导的大尺度操纵、核酸酶介导的大尺度操纵、从头合成大片段DNA以及其他大尺度操纵工具,介绍了它们的基本工作原理与典型应用案例。最后,对大尺度遗传操纵面临的挑战和发展进行了展望。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,de novo 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.

关 键 词：遗传操纵 基因组工程 大片段DNA 酵母 合成生物学 

分 类 号：Q78[生物学—分子生物学]