College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, No. 30 South Puzhu Road, Nanjing, 211816, People's Republic of China.
Jiangsu National Synergetic Innovation Center for Advanced Materials (SICAM), No. 30 South Puzhu Road, Nanjing, 211816, People's Republic of China.
Appl Microbiol Biotechnol. 2019 Apr;103(8):3239-3248. doi: 10.1007/s00253-019-09726-x. Epub 2019 Mar 15.
Microalgae are arguably the most abundant single-celled eukaryotes and are widely distributed in oceans and freshwater lakes. Moreover, microalgae are widely used in biotechnology to produce bioenergy and high-value products such as polyunsaturated fatty acids (PUFAs), bioactive peptides, proteins, antioxidants and so on. In general, genetic editing techniques were adapted to increase the production of microalgal metabolites. The main genome editing tools available today include zinc finger nucleases (ZFNs), transcriptional activator-like effector nucleases (TALENs), and the clustered regularly interspaced short palindromic repeats (CRISPR)/Cas nuclease system. Due to its high genome editing efficiency, the CRISPR/Cas system is emerging as the most important genome editing method. In this review, we summarized the available literature on the application of CRISPR/Cas in microalgal genetic engineering, including transformation methods, strategies for the expression of Cas9 and sgRNA, the CRISPR/Cas9-mediated gene knock-in/knock-out strategies, and CRISPR interference expression modification strategies.
微藻被认为是最丰富的单细胞真核生物，广泛分布于海洋和淡水湖泊中。此外，微藻在生物技术中被广泛用于生产生物能源和高价值产品，如多不饱和脂肪酸（PUFAs）、生物活性肽、蛋白质、抗氧化剂等。一般来说，遗传编辑技术被用于提高微藻代谢产物的产量。目前可用的主要基因组编辑工具包括锌指核酸酶（ZFNs）、转录激活因子样效应物核酸酶（TALENs）和成簇规律间隔短回文重复（CRISPR）/Cas 核酸酶系统。由于其高效的基因组编辑效率，CRISPR/Cas 系统正在成为最重要的基因组编辑方法。在这篇综述中，我们总结了 CRISPR/Cas 在微藻基因工程中的应用的现有文献，包括转化方法、Cas9 和 sgRNA 的表达策略、CRISPR/Cas9 介导的基因敲入/敲除策略以及 CRISPR 干扰表达修饰策略。
