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[Samantha]

Dear all,

i tried to use a LNA miRNA inhibitor (20nM-50nM) to knock down my miRNA of interest by silentfect (BioRad) in a breast cancer cell line ZR75. I got inconsistent experimental results, one time successfully knocked down the miRNA, twice failed with some induction of the miRNA. The knockdown efficiency was checked by TaqMan real time PCR.

1. Has anyone here successfully transfected ZR75? Actually, I have tried many transfection reagents (for both overexpression or knockdown), but couldnt succeed. What reagents did you use? Any suggestions on transfection of this cell line?

2. Has anyone entercoutered induction of miRNA expression when you try to knock down the miRNA?  Any clues to explain this? How did you solve this eventually if you had had this problem before?

Thank you.

Samantha