Cytoplasmic granulation after siRNA transfection - (Mar/10/2007 )
Hello,
After transfecting some cell lines with siRNA (100nM) I see the appearance of cytoplasmic bubbles or granules (by light microscopy) in the perinuclear region of the cell. The majority of the cells are not apoptotic and appear to respond normally to various stimuli but I don't know what these unknown bubbles are. What I do know is that they appear regardless of control non targeting or transcript specific targeting siRNA so it is possible that the transfection reagent (DharmaFect 1) could be causing this phenomena. Does anyone have any idea what this is or what may be causing it? Thanks.
I think it is the stress that the transfection reagent induced to the treated cells. Can you reduce the dosages for siRNA and the transfection reagent and see if the number of such vacules reduces?
After transfecting some cell lines with siRNA (100nM) I see the appearance of cytoplasmic bubbles or granules (by light microscopy) in the perinuclear region of the cell. The majority of the cells are not apoptotic and appear to respond normally to various stimuli but I don't know what these unknown bubbles are. What I do know is that they appear regardless of control non targeting or transcript specific targeting siRNA so it is possible that the transfection reagent (DharmaFect 1) could be causing this phenomena. Does anyone have any idea what this is or what may be causing it? Thanks.
there is a theory that these vacuoles are used by the cell to get rid off toxic agents such as transfection reagents, antibiotics and so on; in some cases depending on your cell type cells stop dividing if strongly vacuolized; I do not think that these vacuoles are the reason but may be regarded as a general feature for toxic stress
thank you both for your comments. if this is a common enough response to siRNA transfection, have any of these theories been published or is there any more information available on these cytoplasmic vacuoles? thanks.