Dear all,
I want to knock down one protein by 50% in HCC cell lines. I want to know whether it is possible and how to achieve it?
Thanks sincerely for your answer.
Zhou.
By using protein depletion method, you may achieve this. It is Auxin-inducible degron (AID) method. You may titrate Auxin concentration and find the right dose for 50% depletion.
The question is for how long one wants the 50% levels to be maintained in the cell in order to examine the effect. I wander if this is OK for long term assays. Alternative ways is heterozygous Knock out.
You could try a transient knockdown of your gene of interest via siRNA or shRNA and see if you can achieve a 50% knockdown of the resulting gene product. It should be possible to get a 50% knockdown via siRNA by playing around with the siRNA concentration you use. But then again this is only transient so the effect will only last for a finite period of time.
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