2 replies to this topic

[Alsulaiman]

Hi folks,

Hope you are well all !!

I did knockdown lately on cell line expressed in GOI and it was tested by qPCR. The results showed that i got moderate suppression in mRNA level in my GOI, following the knockdown i did cell proliferation ( cell viability ) and the results showed that there is an effect on proliferation rate.

]My question is how was an effect on proliferation when there was little knockdown ?


Many thank

Abdullah  

Edited by Alsulaiman, 24 July 2012 - 09:47 PM.

[Jon Moulton]

Assuming the change is proliferation is not due to the moderate knockdown of your intended target, then perhaps it is the interaction of your knockdown reagent with an unintended RNA (an off-target knockdown) that is causing the change in proliferation.

[zodiac1505]

I agree with the earlier post. It could also be that your GOI is upstream of something else and the moderate reduction is amplified downstream leading to the phenotype that you observed. If you are sure that it can't be due to the knockdown, then the most probable reason is off target effect.