Exosomes-mediated synthetic dicer substrates delivery for intracellular dicer imaging detection

In this study, we design a fluorescent labeling Dicer substrate and effectively deliver it into cell by exosomes derived from the target parent cells for intracellular Dicer expression level monitor and gene therapy. Using pre-miRNA let-7a as a model, the Dicer substrates with two terminals labeled with fluorescent and quencher group respectively was obtained by T4 RNA mediated ligase reaction from two short RNA sequences. Then, the substrate was packaged into exosomes by electroporation and delivered to target cells for intracellular dicer imaging detection. After packaging substrates into exosomes with little immunogenicity and good innate biocompatibility by electroporation and delivered to target cells, the Dicer mediated substrate cleavage was effectively monitored by the fluorescence recovery, providing a powerful tool for Dicer analysis. Importantly, the cleaved product exhibited significant suppression toward tumor cell growth and regulated cancer cells cycle. This work might open a new avenue for Dicer analysis and Dicer-related clinical application.Graphical abstractA simple strategy is designed for intracellular Dicer expression level monitor and gene therapy. The synthetic Dicer substrates can cleave in cytoplasm result in fluorescent recovery for Dicer analysis by RNase Dicer through exosome mediated delivery. Another cleaved product with intrinsic gene silence ability trigger significant cancer cells growth suppression and G1/S block.

Source: Biosensors and Bioelectronics - December 10, 2019 Category: Biotechnology Source Type: research