Key Laboratory of Synthetic Biology, CAS Center for Excellence in Molecular Plant Sciences, Institute of Plant Physiology and Ecology, Chinese Academy of Sciences, Shanghai 200032, China.
Key Laboratory of Synthetic Biology, CAS Center for Excellence in Molecular Plant Sciences, Institute of Plant Physiology and Ecology, Chinese Academy of Sciences, Shanghai 200032, China; School of Life Sciences, Henan University, Kaifeng 475004, China.
Biotechnol Adv. 2019 Sep-Oct;37(5):730-745. doi: 10.1016/j.biotechadv.2019.04.002. Epub 2019 Apr 3.
Industrial biotechnology is reliant on native pathway engineering or foreign pathway introduction for efficient biosynthesis of target products. Chromosomal integration, with intrinsic genetic stability, is an indispensable step for reliable expression of homologous or heterologous genes and pathways in large-scale and long-term fermentation. With advances in synthetic biology and CRISPR-based genome editing approaches, a wide variety of novel enabling technologies have been developed for single-step, markerless, multi-locus genomic integration of large biochemical pathways, which significantly facilitate microbial overproduction of chemicals, pharmaceuticals and other value-added biomolecules. Notably, the newly discovered homology-mediated end joining strategy could be widely applicable for high-efficiency genomic integration in a number of homologous recombination-deficient microbes. In this review, we explore the fundamental principles and characteristics of genomic integration, and highlight the development and applications of targeted integration approaches in the three representative industrial microbial systems, including Escherichia coli, actinomycetes and yeasts.
工业生物技术依赖于天然途径工程或外源途径引入，以高效生物合成目标产物。染色体整合具有内在的遗传稳定性，是在大规模和长期发酵中可靠表达同源或异源基因和途径的不可或缺的步骤。随着合成生物学和基于 CRISPR 的基因组编辑方法的进步，已经开发出了各种各样的新型使能技术，可用于一步法、无标记、多基因座生化途径的基因组整合，这极大地促进了微生物对化学品、药物和其他有价值的生物分子的过量生产。值得注意的是，新发现的同源介导的末端连接策略可能广泛适用于许多同源重组缺陷型微生物中的高效基因组整合。在这篇综述中，我们探讨了基因组整合的基本原则和特点，并重点介绍了靶向整合方法在三种代表性工业微生物系统（大肠杆菌、放线菌和酵母）中的开发和应用。
