Hi Scientist,
I am transfecting an oncogenic miRNA into a breast cancer cell line and based on your experience in the mammalian cell line, will such transfection affect the cell morphology?
It is difficult to predict an outcome of any transfection experiment involving miRNA in a cell line. If one includes controls- cells mock transfected, transfected without miRNA and with miRNA, it would be possible to determine whether the miRNA in question would have any effect on the cell morphology post-transfection.
It's not always so straight that transfection will change the morphology. I will suggest add transfection control along with what divakar sir suggested. Transfection Reagent is well known culprit which affects cells.
Hi,
In our previous laboratory, we detected that miRNA transfection with Lipofectamine™ 2000 affected on morphology of MDA-MB-231 breast cancer cell line however, it did not influence on MCF-7, T47D, and MCF-10A cell lines morphology. Therefore, I think it depends to cell type.
Regards,
Yes, I was pointing out this only, Lipofectamine which always a doubtful for me. Means when I started work with miRNA, first of all, Lipofectamine is not at all able to transfect miRNA mimic/inhibitors to cells. I pissed off with that. In complementary effect, cells death & morphology change. I really get surprised how other people does transfection using Lipofectamine. I change the transfection reagent after struggling for 6 month and after understanding the cause. Now, I am using INTERferin from polyplus which works wonderfully. So, I always recommends that.
yes, if say this to Thermo company, then they will suggest to use
Lipofectamine RNAiMAX Reagent.
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