在真核细胞中分子的转运是一个动态过程，Cyto-Tracer提供一种能力，照亮细胞的组成部分并监视其活动，同时定位细胞器，并跟踪细胞内吞作用和细胞外吐作用。SBI创造了一条基于慢病毒载体的Cyto-Tracer应用于长期和深入的实验，其中运用GFP融合蛋白来标记细胞组件，细胞器，小泡和架构。Cyto-Tracer可以用于转染，以及包装进病毒以在原代细胞、肿瘤细胞和干细胞中生产出稳定的GFP跟踪细胞系。
亮点：
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基于慢病毒的细胞跟踪系统
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对与别的蛋白质进行共同定位的研究很理想
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实时监控蛋白质的转运
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有为细胞器、小泡和其它组分构建的质粒
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明亮的对光稳定的GFP对细胞没有干扰作用。
Cyto-Tracer：pCT-CD63-GFP
靶定位：Exosome/Secretory
标记物靶肽：CD63 Tetraspanin
Supporting Data
See some of our exosome Cyto-Tracers in action
The following figure and videos are from:
Garcia NA, et al. Glucose Starvation in Cardiomyocytes Enhances Exosome Secretion and Promotes Angiogenesis in Endothelial Cells. PLoS ONE. 2015. 10(9). PMCID: PMC4578916.
Figure 4 from Garcia, et al. Exosome transfer from CMs to ECs. (A) H9c2 transfected with pCT-CD63-GFP. FACS analysis of 90% GFP positive cells. (B) Representative images from confocal time-lapse microscopy of mouse ECs (ACTB-DsRed) (EC; red) co-cultured with H9C2-CD63-GFP cells (green), previously cultured for 24 h in +/-St medium. Exosome transfer from H9C2 CMs to EC can be observed (S1 and S2 Movies, below). White arrows show CD63-GFP structures inside ECs (C) Representative immunostaining of H9C2-CD63-GFP and HUVEC co-cultures; anti-GFP (green) and anti-CD31 (red). The images illustrate GFP fluorescence from CD63-GFP exosomes in CD31-positive cells (red) after 24 h incubation in +/-St medium. White arrows show CD63-GFP structures inside ECs.
Video S1 from Garcia, et al. Exosome transfer from H9C2-CD63-GFP (green) to endothelial DsRed cells (red) under +St conditions.
Video S2 from Garcia, et al. Exosome transfer from H9C2-CD63-GFP (green) to endothelial DsRed cells (red) under -St conditions.
Labeled exosomes. (Top panels) CD63-GFP Cyto-Tracers transfected into a human fibrosarcoma cell line. (Bottom panels) CD9-GFP and CD9-RFP Cyto-Tracers co-transfected into HEK293 cells.
