Pease, we are new to the field, and text in manuscripts are sometimes confusing. I want to study the effect of overexpression and down expression of a given miRNA in tumor development. Is it enough to transfect miRNAS or anit-miRNAs into a cell line and inject them in nude mice? Or do I have to generate a stable cell line expressing that miRNA (shRNA) or its anit-miRNA and then inject these cells into nude mice? I read some papers where they just say mir transfected cells were injected into nude mice but neither in the methods or other parts of the paper there are better explanations.
The effect of transient transfection lasts 3-4 days. So it is pretty clear that you can not perform a proper in vivo experiment using transiently transfected cells. You can either administrate miRNA mimics directly to the mice, but it will be a "whole body" effect. Otherwise you have to generate a stable cell line.
It depends on the type of experiment that you want to performe. If you want to do an extravasation assay (it lasts 48h) , you can transfect your cells with pre-miRNA or anti-miRNA. If you want to see metastatization from primary tumor or from tail vein injection or check primary tumor growth, you have to use a stable cell line that overexpress your miRNA of interess or downmodulate it (you can use the sponges).
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