基于代谢工程与生物正交反应的病毒原位标记技术研究

Research on Virus in Situ Labeling Technology Based on Metabolic Engineering and Bioorthogonal Reaction

示踪病毒在细胞内的感染路径对探究病毒的感染机制具有重要意义,病毒标记是实现病毒示踪的关键。针对目前病毒标记方法中存在的不足,该文提出了一种基于代谢工程的病毒原位生物正交标记技术。脂质/氨基酸/单糖的叠氮衍生物经细胞代谢将叠氮基团修饰在病毒表面,被修饰病毒粒子识别结合宿主细胞后与携带二苯基环辛炔(DBCO)的荧光探针经生物正交反应连接从而实现病毒标记。结果显示,叠氮基团在囊膜病毒和非囊膜病毒表面都有很好的修饰效果,并且在原位生物正交中携带二苯基环辛炔基团的荧光探针可快速捕捉到叠氮修饰的病毒粒子实现病毒标记。该标记方法不仅操作简单,而且适用于囊膜病毒与非囊膜病毒的标记。

Tracking virus infection pathway in cells is of great significance for understanding the viral infection mechanism,and the successful labeling on viruses is the key to achieve viruses tracking.In this study,we proposed the in situ bioorthogonal strategy based on the bioorthogonal chemistry and metabolic engineering for overcoming defects of traditional virus labeling methods.Azido motifs were incorporated into virions through cell metabolism with lipid/amino acid/monosaccharide azide derivatives,and the modified virions were in situ labeled with dibenzocyclooctyne-functionalized fluorescent probes through the bioorthogonal reaction.The results show that viruses incorporated with azide motifs could be quickly and effectively captured by fluorescent probes to achieve viruses in situ labeling.Significantly,the in situ bioorthogonal labeling strategy is a general method for enveloped and non-enveloped viruses labeling with simple operations.