Liu Zihe, Liang Youyun, Ang Ee Lui, Zhao Huimin
Metabolic Engineering Research Laboratory, Science and Engineering Institutes, Agency for Science, Technology and Research , Singapore.
Department of Chemical and Biomolecular Engineering, University of Illinois at Urbana-Champaign , Urbana, Illinois 61801, United States.
ACS Synth Biol. 2017 Apr 21;6(4):601-609. doi: 10.1021/acssynbio.6b00331. Epub 2017 Jan 24.
Genome integration is a powerful tool in both basic and applied biological research. However, traditional genome integration, which is typically mediated by homologous recombination, has been constrained by low efficiencies and limited host range. In recent years, the emergence of homing endonucleases and programmable nucleases has greatly enhanced integration efficiencies and allowed alternative integration mechanisms such as nonhomologous end joining and microhomology-mediated end joining, enabling integration in hosts deficient in homologous recombination. In this review, we will highlight recent advances and breakthroughs in genome integration methods made possible by programmable nucleases, and their new applications in synthetic biology and metabolic engineering.
基因组整合在基础生物学研究和应用生物学研究中都是一种强大的工具。然而,传统的基因组整合通常由同源重组介导,其效率低下且宿主范围有限。近年来,归巢内切酶和可编程核酸酶的出现极大地提高了整合效率,并允许诸如非同源末端连接和微同源性介导的末端连接等替代整合机制,从而能够在缺乏同源重组的宿主中进行整合。在这篇综述中,我们将重点介绍可编程核酸酶实现的基因组整合方法的最新进展和突破,以及它们在合成生物学和代谢工程中的新应用。