新型基因编辑技术加速盐藻表达系统的成熟和应用  

Novel Gene Editing Technology Accelerates Maturation and Application of Dunaliella salina Expression Systems

作　　者：黄晓雯 皇甫雨静 姬聪浩 代月优 李姝璇 冯书营 HUANG Xiaowen;HUANGFU Yujing;JI Conghao;DAI Yueyou;LI Shuxuan;FENG Shuying(Henan University of Chinese Medicine,Medical College,Zhengzhou 450046,China;Henan Engineering Research Center of Special Medical Food of Traditional Chinese Medicine,Zhengzhou 450046,China;Henan Engineering Research Center of Medicinal and Food Homologous Traditional Chinese Medicine,Zhengzhou 450046,China)

机构地区：[1]河南中医药大学医学院,郑州450046 [2]河南省中医药特医食品工程研究中心,郑州450046 [3]河南省药食同源中药工程研究中心,郑州450046

出　　处：《中国生物工程杂志》2024年第4期102-111,共10页China Biotechnology

基　　金：国家自然科学基金(U1804112);河南省自然科学基金(232300421164);河南省高等学校重点科研项目计划基础研究专项(23ZX005)资助项目。

摘　　要：盐藻因具备低成本、易培养、便于转基因操作、可调控的转录和翻译等优点,已成为基因工程领域备受关注的新型表达系统。但盐藻表达系统存在低表达和不稳定遗传等技术瓶颈,严重限制了其应用。CRISPR/Cas系统凭借高稳定性、高靶点特异性、可编程性等优点,可作为助推盐藻表达系统改进和应用的重要工具,从而大幅提高盐藻表达系统重组蛋白、脂质积累和胡萝卜素等高附加值产物的产量。由于盐藻兼具原核细胞和真核细胞的双重特点,基因编辑存在较大的难度和限制性,因而必须不断提高基因编辑技术在盐藻中的精确性和编辑效率,以确保目标基因得到准确修饰。探讨盐藻表达系统的基因工程研究现状,包括盐藻转化方法、外源基因的表达和基因编辑的研究现状等,分析CRISPR/Cas编辑技术在盐藻中面临的主要挑战,提出未来的发展方向和增效策略,以期加速盐藻表达系统的成熟和应用。Dunaliella salina(D.salina)has become a promising expression system in genetic engineering due to its low cost,easy cultivation,simple transgenic operation,and controllable transcription and translation characteristics.However,there are technical bottlenecks such as low expression and unstable inheritance in the D.salina expression system,which severely limit its application.With the advantages of high stability,the high target specificity and programmability,the CRISPR/Cas system can be used as an important tool to promote the improvement and application of the D.salina expression system,thereby greatly increasing the production of high value-added products such as recombinant proteins,lipid accumulation and carotene.Due to the dual characteristics of prokaryotic and eukaryotic cells,salt algae have greater difficulties and limitations in the gene editing process.Therefore,it is necessary to continuously improve the accuracy and editing efficiency of gene editing technology in D.salina to ensure that the target genes are accurately modified.On this basis,the advantages of genetic engineering of D.salina expression system were comprehensively reviewed,including the transformation methods,exogenous gene expression and the current research status of gene editing.Meanwhile,the main challenges of CRISPR/Cas editing technology in D.salina are highlighted,the future development direction and improvement strategy are proposed,and the application prospect of this technology is envisioned to accelerate the early maturation and popularization of D.salina expression system.

关 键 词：盐藻 新型表达系统 基因编辑 CRISPR/Cas9系统 

分 类 号：Q819[生物学—生物工程]