I am trying to knockdown gene of my interest in U251 and U87MG cell lines I found that after siRNA transfection my gene of interest showing upregulation upto 2 folds even after several repetitions. I am using siRNA pool from GE for silencing. Has anyone faced this? kindly help me with the troubleshooting
HI,
excluding technical issues (could be countless), which is you gene of interest? Have you checked if there are pseudogenes of your target gene? Maybe your gene upregulation might be caused as a consequence of gene-pseudogene competition for siRNA.
Hi preethi
siRNA will block the protein production by blocking the RNA. do you have a feedback control of the protein amount control, inducing a higher production of the RNA, even if it's blocked??
fred
There are can be two reasons:
1. Technical Experimental Issues (you need to repeat the experiment and see if the results are consistent)
2. Existence of feed-back loops between the gene and other genes.
Hello all,
Thank you very much for your feedback, I found there was some experimental error and rectified now I repeated thrice now it is working fine
@Svetlana,
No I did not compare it with the scrambled control but with untreated.
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