Scientific reports, 2022-01, Vol.12 (1), p.496-496, Article 496
2022
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- Autor(en) / Beteiligte
- Titel
- A new transgene mouse model using an extravesicular EGFP tag enables affinity isolation of cell-specific extracellular vesicles
- Ist Teil von
- Scientific reports, 2022-01, Vol.12 (1), p.496-496, Article 496
- Ort / Verlag
- England: Nature Publishing Group
- Erscheinungsjahr
- 2022
- Quelle
- Free E-Journal (出版社公開部分のみ）
- Beschreibungen/Notizen
- The in vivo function of cell-derived extracellular vesicles (EVs) is challenging to establish since cell-specific EVs are difficult to isolate and differentiate. We, therefore, created an EV reporter using truncated CD9 to display enhanced green fluorescent protein (EGFP) on the EV surface. CD9truc-EGFP expression in cells did not affect EV size and concentration but enabled co-precipitation of EV markers TSG101 and ALIX from the cell-conditioned medium by anti-GFP immunoprecipitation. We then created a transgenic mouse where CD9truc-EGFP was inserted in the inverse orientation and double-floxed, ensuring irreversible Cre recombinase-dependent EV reporter expression. We crossed the EV reporter mice with mice expressing Cre ubiquitously (CMV-Cre), in cardiomyocytes (αMHC-MerCreMer) and renal tubular epithelial cells (Pax8-Cre), respectively. The CD9truc-EGFP positive mice showed Cre-dependent EGFP expression, and plasma CD9truc-EGFP EVs were immunoprecipitated only from CD9truc-EGFP positive CD9truc-EGFPxCMV-Cre and CD9truc-EGFPxαMHC-Cre mice, but not in CD9truc-EGFPxPax8-Cre and CD9truc-EGFP negative mice. In urine samples, CD9truc-EGFP EVs were detected by immunoprecipitation only in CD9truc-EGFP positive CD9truc-EGFPxCMV-Cre and CD9truc-EGFPxPax8-Cre mice, but not CD9truc-EGFPxαMHC-Cre and CD9truc-EGFP negative mice. In conclusion, our EV reporter mouse model enables Cre-dependent EV labeling, providing a new approach to studying cell-specific EVs in vivo and gaining a unique insight into their physiological and pathophysiological function.
- Sprache
- Englisch
- Identifikatoren
- ISSN: 2045-2322eISSN: 2045-2322DOI: 10.1038/s41598-021-04512-0Titel-ID: cdi_doaj_primary_oai_doaj_org_article_fb27a2cf4445455e9edf68103b42b4e3
- Format
- –
- Schlagworte
- Animals, Cardiomyocytes, CD9 antigen, Cell surface, Chromatography, Cre recombinase, Epithelial cells, Epithelial Cells - metabolism, Extracellular vesicles, Extracellular Vesicles - chemistry, Extracellular Vesicles - genetics, Extracellular Vesicles - metabolism, Genes, Reporter, Green fluorescent protein, Green Fluorescent Proteins - chemistry, Green Fluorescent Proteins - genetics, Green Fluorescent Proteins - metabolism, Immunoprecipitation, Kidney Tubules, Distal - cytology, Kidney Tubules, Distal - metabolism, Labeling, Mice, Mice, Transgenic - genetics, Mice, Transgenic - metabolism, Molecular weight, Myocytes, Cardiac - metabolism, Organ Specificity, Pax8 protein, Physiology, Plasma, Proteins, Transgenes, Transgenic mice