Hi. I am studying the role of exosomes in cancer metastasis. Exosomes derived from cultured tumor cells could be isolated and was identified by various methods. I was wondering if it would be possible to increase or decrease a specific microRNA inside the isolated exosomes in vitro? The plan is to use the microRNA modified exosomes to investigate the downstream effects or mechanisms if that could be done. Thanks a lot.
I'm skeptical about knocking out microRNA in exosomes. Instead, you can KO the miRNA in the tumor cell and the isolate the exosomes, run pcr to ensure the absence of the particular miRNA and proceed with the downstream analysis.
As Jayabalan suggested you can KO a particular miRNA in the original cells to get exosomes with less concentration of this miRNA (we have already published that)
https://www.ncbi.nlm.nih.gov/pubmed/27650731
On the other hand there are some hypothesis suggesting that decreasing the expression of the mRNA target of the miRNA in the cells that release the exosomes you could manipulate the miRNA cargo of them.
Hi Xianchong,
Outside of directly transfecting the exosomes (exofect) with your miRNA, there are some engineering options that may interest you that allow you to increase the amount of a specific miRNA that gets packaged into exosomes. The XMIR technology is a plasmid that you can transfect your cells that will cause all secreted exosomes to express your desired miRNA with higher efficiency. This is possible via an xmir tag which is recognized by cellular proteins which then co-localize to the membrane to be incorporated in exosome biogenesis.
Best of luck.
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