外泌体(Exosome)是由细胞分泌而来的微小囊泡,直径约为 30-150 nm,天然存在于血液、尿液、唾液、母乳和细胞培养基等生物体液中,几乎所有类型的细胞都可以产生并释放外泌体。目前,外泌体因其在细胞间通讯中的作用而被广泛地认识到,比如将不同生物分子(包括蛋白质、RNA和脂质)从一个细胞传递到另一个细胞。外泌体具有共同的特征,如某些四跨膜蛋白超家族(CD9、CD63和CD81)、热休克蛋白、生物发生相关蛋白、膜转运和融合蛋白、核酸(mRNA、miRNA、长非编码RNA和DNA)和脂质。
外泌体的这些特性为诊断和治疗提供了新的思路。细胞间囊泡运输在人类健康和疾病相关的许多方面发挥着重要作用,包括发育、免疫、组织稳态、癌症和神经退行性疾病等。因此,外泌体作为生物标志物在临床上的应用、以及治疗的用途越来越受到人们的关注。
Genecopoeia提供携带不同标记(tdTomato, mCherry-Flag, Flag-Gluc)的外泌体标志物CD9, CD63和CD81的预制慢病毒。可用于示踪外泌体的形成、分泌、靶向和运输机制。
另外,Genecopoeia还提供可用于动物活体实验研究的外泌体预制慢病毒,以及外泌体稳转细胞株构建服务。有意请联系我们,致电4006-020-200,或发送邮件至sales@igenebio.com。
图1: CD63外泌体标志物蛋白印迹。在293T细胞中分别表达A, B, C, D质粒。阳性质粒是Flag-EGFP。蛋白印迹样品均来源于293T细胞上清并经过纯化。左图使用1:1000 anti-CD63 抗体和1:5000 anti-mouse IgG抗体进行检测,右图使用1:1000 anti-Flag抗体和1: 5000 anti-mouse IgG抗体进行检测。相关蛋白的预期大小分别为:Flag: 24 kDa, Gluc: 18 kDa, Nluc: 19 kDa, mCherry: 26 kDa, CD63: 53 kDa, Flag-Gluc-CD63: 73 kDa, Flag-mCherry-CD63: 81 kDa。
图2:FACS分析被标记的外泌体的细胞培养实验。在HEK293T细胞系中表达CD63-tdTomato和CD81-tdTomato融合蛋白。从细胞上清液中分离外泌体,用于细胞培养实验。将新铺板的HEK293T细胞(未处理)与分离的外泌体一起孵育6小时,然后进行FACS分选分析。
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