🧫 Experiment Protocol
ValidationCD9Transgenic TIGER miceproposed
A transgenic mouse line was generated containing a CRE-recombinase inducible CAG promoter driving CD9 protein fused to Turbo-GFP (TIGER mouse). The mice were characterized by electroporation with CAG-CRE plasmids or crossing with tamoxifen inducible CAG-CRE-ERT2 or nestin-CRE-ERT2 mice to achieve cell-type specific labeling of extracellular vesicles. CD9-GFP labeled cells were identified as glutamine synthetase and glial fibrillary acidic protein positive astrocytes. This experiment established a novel tool for studying EV biogenesis, release, and cellular uptake in vivo with cell-type specificity.
PRIMARY OUTCOME
Generation of functional CD9-GFP reporter mice for EV tracking
EXPECTED OUTCOMES
Cell-type specific labeling of EVs with CD9-GFP in astrocytes
SUCCESS CRITERIA
Successful CD9-GFP expression in target cells and EV labeling
PROTOCOL
Transgenic mouse generation with CRE-inducible CD9-Turbo-GFP construct, electroporation with CAG-CRE plasmids, crossing with CRE driver lines, immunofluorescence characterization