Wu Yijian, Feng Sai, Sun Zeao, Hu Yan, Jia Xiao, Zeng Bin
College of Pharmacy, Shenzhen Technology University, Shenzhen, Guangdong, China.
Front Bioeng Biotechnol. 2023 Oct 5;11:1249841. doi: 10.3389/fbioe.2023.1249841. eCollection 2023.
is one of the most extensively used biosynthetic systems for the production of diverse bioproducts, especially biotherapeutics and recombinant proteins. Because the expression and insertion of foreign genes are always impaired by the endogenous factors of and nonproductive procedures, various technologies have been developed to enhance the strength and efficiency of transcription and facilitate gene editing procedures. Thus, the limitations that block heterologous protein secretion have been overcome. Highly efficient promoters responsible for the initiation of transcription and the accurate regulation of expression have been developed that can be precisely regulated with synthetic promoters and double promoter expression systems. Appropriate codon optimization and harmonization for adaption to the genomic codon abundance of are expected to further improve the transcription and translation efficiency. Efficient and accurate translocation can be achieved by fusing a specifically designed signal peptide to an upstream foreign gene to facilitate the secretion of newly synthesized proteins. In addition to the widely applied promoter engineering technology and the clear mechanism of the endoplasmic reticulum secretory pathway, the innovative genome editing technique CRISPR/Cas (clustered regularly interspaced short palindromic repeats/CRISPR-associated system) and its derivative tools allow for more precise and efficient gene disruption, site-directed mutation, and foreign gene insertion. This review focuses on sophisticated engineering techniques and emerging genetic technologies developed for the accurate metabolic regulation of the expression system.
是用于生产多种生物制品，特别是生物治疗药物和重组蛋白的最广泛使用的生物合成系统之一。由于外源基因的表达和插入总是受到内源性因素和非生产性程序的影响，因此已经开发了各种技术来增强转录的强度和效率，并促进基因编辑程序。因此，阻碍异源蛋白分泌的限制已被克服。已经开发出负责转录起始和精确表达调控的高效启动子，其可以通过合成启动子和双启动子表达系统进行精确调控。为适应基因组密码子丰度而进行的适当密码子优化和协调有望进一步提高转录和翻译效率。通过将专门设计的信号肽与上游外源基因融合，可以实现高效准确的转运，以促进新合成蛋白质的分泌。除了广泛应用的启动子工程技术和内质网分泌途径的明确机制外，创新的基因组编辑技术CRISPR/Cas（成簇规律间隔短回文重复序列/CRISPR相关系统）及其衍生工具允许更精确和高效的基因破坏、定点突变和外源基因插入。本综述重点介绍为精确代谢调控表达系统而开发的复杂工程技术和新兴遗传技术。
