To inhibit a specific enzyme inside a cell, and study its role in a cell or in vivo, the first problem seems valid is that does the inhibitor just inhibit the enzyme you want or a number of other enzyme that you don't know?
Is there any solution? Is it possible to use antibody or such things to do that?
Thank you
Dear Azad
First, look for a specific inhibitor of the enzyme. This inhibitor should be able to cross the cell membrane and soluble in physiologic cionditions. You may varify its specificity in vitro, which might be laborous. Antibodies will not cross the cell membrane. You have to permeabilize the cells which will kill them. If there is no secure and specific inhibitor, I would try to knock down the enzyme by siRNA or CRISP/Cas9.
best wishes
Gernot
Hello,
Wich enzyme do you want to inhibit ? Maybe you can found a pharmacological inhibitor. If you want a very specific inhibition, the best way is a siRNA, and if it doesn't work, you can try a CRISPR/cas9 experiment which is very efficient but very long and expensive. Don't forget to confirm your inhibition by a Western Blot.
Best regards
Chloé
There are specific inhibitors known for many enzymes. Often these inhibitors can be used in vitro and in vivo depending on how important your enzyme is and the characteristics of the individual inhibitor (like toxicity, etc.).
You have to check literature on your enzyme of interest, chances are someone already identified its specific inhibitors.
As Gernot proposed, if there are no specific inhibitors, there are other ways to inhibit its function in vivo.
Obviously you can in both conditions. The explanations by above are in right direction. Good luck!!!
Hello Azad
I think the best way do it in vivo is the siRNA and in vitro you may try an inhibitor
Good luck
If you know the aminoacids in the active center of the enzyme then you may use a specific inhibitor in vitro or if the enzyme is metaloenzyme then you may use a chelator to inhibit.
It depends on what kind of enzyme you want to block. Might be a popular enzyme like kinases whose specific inhibitor already known.You may also completely block its expression before the respected mRNA of that enzyme is get translated inside the cells using modern gene silencing techniques using siRNA, CRISPR/cas9 etc.
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