Chaoshan Han , Junjie Yang , Tingting Yin , Junqing An , Aijun Qiao , Yangpo Cao , Yuliang Feng , Haocheng Lu , Ying Wang , Liang Yang , Gangjian Qin
{"title":"CD63-snorkel标记法分离外泌体","authors":"Chaoshan Han , Junjie Yang , Tingting Yin , Junqing An , Aijun Qiao , Yangpo Cao , Yuliang Feng , Haocheng Lu , Ying Wang , Liang Yang , Gangjian Qin","doi":"10.1016/j.vesic.2023.100031","DOIUrl":null,"url":null,"abstract":"<div><p>Exosomes (Exo) are important mediators of inter-cellular communications; however, no effective method is available for isolating, thus characterizing, cellular-specific exosomes <em>in vivo</em>. Since CD63 is a reliable marker for exosomes, we have developed a tagging strategy, term “CD63-Snorkel (CD63-SNKL)”, in which CD63 at its intracellular C-terminus was fused to a fragment of PDGFRB that contains the transmembrane domain tethered to multiple epitope tags (HA, His, and FLAG) displayed in tandem on surface. We found that the CD63-SNKL protein has similar subcellular localizations as endogenous CD63 and can be effectively sorted into Exo. Furthermore, Exo secreted from CD63-SNKL–transduced cells can be effectively captured on anti-HA magnetic beads and eluted with HA peptides. Thus, CD63-SNKL may be engineered for isolating and tracking endogenous tissue-specific Exo <em>in vivo</em>.</p></div>","PeriodicalId":73007,"journal":{"name":"Extracellular vesicle","volume":"2 ","pages":"Article 100031"},"PeriodicalIF":0.0000,"publicationDate":"2023-10-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2773041723000100/pdfft?md5=4118b6e1865381af9c561c620692b519&pid=1-s2.0-S2773041723000100-main.pdf","citationCount":"0","resultStr":"{\"title\":\"CD63-snorkel tagging for isolation of exosomes\",\"authors\":\"Chaoshan Han , Junjie Yang , Tingting Yin , Junqing An , Aijun Qiao , Yangpo Cao , Yuliang Feng , Haocheng Lu , Ying Wang , Liang Yang , Gangjian Qin\",\"doi\":\"10.1016/j.vesic.2023.100031\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><p>Exosomes (Exo) are important mediators of inter-cellular communications; however, no effective method is available for isolating, thus characterizing, cellular-specific exosomes <em>in vivo</em>. Since CD63 is a reliable marker for exosomes, we have developed a tagging strategy, term “CD63-Snorkel (CD63-SNKL)”, in which CD63 at its intracellular C-terminus was fused to a fragment of PDGFRB that contains the transmembrane domain tethered to multiple epitope tags (HA, His, and FLAG) displayed in tandem on surface. We found that the CD63-SNKL protein has similar subcellular localizations as endogenous CD63 and can be effectively sorted into Exo. Furthermore, Exo secreted from CD63-SNKL–transduced cells can be effectively captured on anti-HA magnetic beads and eluted with HA peptides. Thus, CD63-SNKL may be engineered for isolating and tracking endogenous tissue-specific Exo <em>in vivo</em>.</p></div>\",\"PeriodicalId\":73007,\"journal\":{\"name\":\"Extracellular vesicle\",\"volume\":\"2 \",\"pages\":\"Article 100031\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2023-10-26\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.sciencedirect.com/science/article/pii/S2773041723000100/pdfft?md5=4118b6e1865381af9c561c620692b519&pid=1-s2.0-S2773041723000100-main.pdf\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Extracellular vesicle\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S2773041723000100\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Extracellular vesicle","FirstCategoryId":"1085","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S2773041723000100","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
Exosomes (Exo) are important mediators of inter-cellular communications; however, no effective method is available for isolating, thus characterizing, cellular-specific exosomes in vivo. Since CD63 is a reliable marker for exosomes, we have developed a tagging strategy, term “CD63-Snorkel (CD63-SNKL)”, in which CD63 at its intracellular C-terminus was fused to a fragment of PDGFRB that contains the transmembrane domain tethered to multiple epitope tags (HA, His, and FLAG) displayed in tandem on surface. We found that the CD63-SNKL protein has similar subcellular localizations as endogenous CD63 and can be effectively sorted into Exo. Furthermore, Exo secreted from CD63-SNKL–transduced cells can be effectively captured on anti-HA magnetic beads and eluted with HA peptides. Thus, CD63-SNKL may be engineered for isolating and tracking endogenous tissue-specific Exo in vivo.
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