Part:BBa_K1633007:Experience

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Applications of BBa_K1633007

USAGE AND BIOLOGY

This part is artificial designed to target and downregulate GFP protein in GFP-transgenic mice or GFP-overexpressed cells.

CHARACTERIZATION

To determine whether siRNA delivered via RVG exosomes can pass through the BBB and regulate endogenous gene expression, we packaged siRNA against green fluorescent protein (GFP) into RVG exosomes and injected them into GFP-transgenic mice through the tail vein. Then, the GFP levels in various tissues were determined by measuring fluorescence emission using a fluorescence microscope. Compared with control mice, injection of the RVG exosomes loaded with GFP siRNA dramatically reduced GFP levels in different parts of the brain of GFP-transgenic mice. In contrast, unmodified exosomes loaded with GFP siRNA did not induce obvious GFP silencing in mouse brain. On the other hand, while unmodified exosomes loaded with GFP siRNA had significant effect on GFP levels in lung, liver and spleen of GFP-transgenic mice, RVG exosomes loaded with GFP siRNA only induced a slight but non-significant GFP silencing in these tissues. The results successfully demonstrate that exosome-packaged siRNA can be delivered to various tissues and thus silence endogenous gene expression. The results also indicate that RVG peptide on the surface of exosome has some selectivity for neuronal tissues, which may simultaneously prevent siRNA from spreading to non-neuronal tissues.

Figure 15. Fluorescence confocal microscopy photographs showing sections from different tissues of GFP-transgenic mice. GFP-transgenic mice were intravenously injected with saline (control) or with GFP siRNA loaded in normal exosomes (siRNA-exosome) or RVG exosomes (siRNA-RVG exosome).

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