Li Mengyuan, Huo Yi-Xin, Guo Shuyuan
Key Laboratory of Molecular Medicine and Biotherapy, School of Life Science, Beijing Institute of Technology, No. 5 South Zhongguancun Street, Beijing 100081, China.
Biology (Basel). 2022 Apr 9;11(4):571. doi: 10.3390/biology11040571.
Nonmodel microbes with unique and diverse metabolisms have become rising stars in synthetic biology; however, the lack of efficient gene engineering techniques still hinders their development. Recently, the use of base editors has emerged as a versatile method for gene engineering in a wide range of organisms including nonmodel microbes. This method is a fusion of impaired CRISPR/Cas9 nuclease and base deaminase, enabling the precise point mutation at the target without inducing homologous recombination. This review updates the latest advancement of base editors in microbes, including the conclusion of all microbes that have been researched by base editors, the introduction of newly developed base editors, and their applications. We provide a list that comprehensively concludes specific applications of BEs in nonmodel microbes, which play important roles in industrial, agricultural, and clinical fields. We also present some microbes in which BEs have not been fully established, in the hope that they are explored further and so that other microbial species can achieve arbitrary base conversions. The current obstacles facing BEs and solutions are put forward. Lastly, the highly efficient BEs and other developed versions for genome-wide reprogramming of cells are discussed, showing great potential for future engineering of nonmodel microbes.
具有独特多样代谢方式的非模式微生物已成为合成生物学领域的后起之秀;然而,缺乏高效的基因工程技术仍然阻碍着它们的发展。最近,碱基编辑器的应用已成为在包括非模式微生物在内的广泛生物体中进行基因工程的一种通用方法。该方法是受损的CRISPR/Cas9核酸酶与碱基脱氨酶的融合,能够在靶位点实现精确的点突变而不诱导同源重组。本综述更新了碱基编辑器在微生物中的最新进展,包括对所有已被碱基编辑器研究过的微生物的总结、新开发的碱基编辑器的介绍及其应用。我们提供了一份全面总结碱基编辑器在非模式微生物中具体应用的列表,这些应用在工业、农业和临床领域发挥着重要作用。我们还介绍了一些碱基编辑器尚未完全建立的微生物,希望对它们进行进一步探索,以便其他微生物物种能够实现任意碱基转换。提出了碱基编辑器目前面临的障碍及解决方案。最后,讨论了用于细胞全基因组重编程的高效碱基编辑器和其他改进版本,展示了其在未来非模式微生物工程中的巨大潜力。