shRNA gene silencing Human - Islets of langerhans

shRNA is a part of RNA interference (RNAi) mechanism where an artificial RNA molecule with a tight hairpin turn could be introduced in cells by delivery of plasmids or through viral or bacterial vectors. Sometimes you see that the shRNA transfection was successful but could not show the protein knockdown with western blot. In that case, check knockdown efficiency on RNA level first by qPCR. If it doesn't work, check the protein level at 72 hours or later. This will ensure that the utilization of existing protein and the "real" effect of mRNA downregulation on the protein level. You can also use more than 3 shRNA targeting sequences for one gene. In addition, using a lentivirus system will greatly help to enhance shRNA knockdown efficiency.

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