化学
外体
核酸
DNA
末端脱氧核苷酸转移酶
微泡
生物化学
细胞生物学
基因
生物
小RNA
细胞凋亡
标记法
作者
Wenjiao Fan,Pihua Han,Qinya Feng,Yuanyuan Sun,Wei Ren,Thomas S. Lawson,Chenghui Liu
标识
DOI:10.1021/acs.analchem.1c04636
摘要
As generally acknowledged, terminal deoxynucleotidyl transferase (TdT) can only elongate DNA substrates from their 3'-OH ends. Herein, for the first time, we report that TdT-catalyzed DNA polymerization can directly proceed on the exosome membrane without the mediation of any nucleic acids. We prove that both the glycosyl and phenolic hydroxyl groups on the membrane proteins can initiate the DNA polymerization. Accordingly, we have developed powerful strategies for high-sensitive exosome profiling based on a conventional flow cytometer and an emerging CRISPR/Cas system. By using our strategy, the featured membrane protein distributions of different cancer cell-derived exosomes can be figured out, which can clearly distinguish plasma samples of breast cancer patients from those of healthy people. This work paves new ways for exosome profiling and liquid biopsy and expands the understanding of TdT, holding great significance in developing TdT-based sensing systems as well as establishing protein/nucleic acid hybrid biomaterials.
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