Ma Ke, Deng Lei, Wu Haizhen, Fan Jianhua
State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, 130 Meilong Road, Shanghai, 200237, People's Republic of China.
Department of Applied Biology, East China University of Science and Technology, Shanghai, 200237, People's Republic of China.
Bioresour Bioprocess. 2022 Aug 13;9(1):83. doi: 10.1186/s40643-022-00568-6.
Microalgae are cosmopolitan organisms in nature with short life cycles, playing a tremendous role in reducing the pressure of industrial carbon emissions. Besides, microalgae have the unique advantages of being photoautotrophic and harboring both prokaryotic and eukaryotic expression systems, becoming a popular host for recombinant proteins. Currently, numerous advanced molecular tools related to microalgal transgenesis have been explored and established, especially for the model species Chlamydomonas reinhardtii (C. reinhardtii hereafter). The development of genetic tools and the emergence of new strategies further increase the feasibility of developing C. reinhardtii chloroplasts as green factories, and the strong genetic operability of C. reinhardtii endows it with enormous potential as a synthetic biology platform. At present, C. reinhardtii chloroplasts could successfully produce plenty of recombinant proteins, including antigens, antibodies, antimicrobial peptides, protein hormones and enzymes. However, additional techniques and toolkits for chloroplasts need to be developed to achieve efficient and markerless editing of plastid genomes. Mining novel genetic elements and selectable markers will be more intensively studied in the future, and more factors affecting protein expression are urged to be explored. This review focuses on the latest technological progress of selectable markers for Chlamydomonas chloroplast genetic engineering and the factors that affect the efficiency of chloroplast protein expression. Furthermore, urgent challenges and prospects for future development are pointed out.
微藻是自然界中广泛分布的生物，生命周期短，在减轻工业碳排放压力方面发挥着巨大作用。此外，微藻具有光合自养以及拥有原核和真核表达系统的独特优势，成为重组蛋白的热门宿主。目前，已经探索并建立了许多与微藻转基因相关的先进分子工具，特别是针对模式物种莱茵衣藻（以下简称衣藻）。遗传工具的发展和新策略的出现进一步提高了将衣藻叶绿体开发为绿色工厂的可行性，衣藻强大的遗传可操作性使其作为合成生物学平台具有巨大潜力。目前，衣藻叶绿体能够成功生产大量重组蛋白，包括抗原、抗体、抗菌肽、蛋白质激素和酶。然而，需要开发更多用于叶绿体的技术和工具包，以实现质体基因组的高效无标记编辑。挖掘新的遗传元件和选择标记将在未来得到更深入的研究，并且迫切需要探索更多影响蛋白质表达的因素。本文综述聚焦于衣藻叶绿体基因工程选择标记的最新技术进展以及影响叶绿体蛋白表达效率的因素。此外，还指出了未来发展面临的紧迫挑战和前景。
