Hi Everyone,
I'm trying to transfect ear cell lines with gene-specific siRNA and also expression plasmid, and then treat them with a drug to see the difference in response. Transfecting 24h before treatment did not change cell response at all.
Based on other papers, that drugs changes the expression of my target gene within 6 hours. So I was wondering is it a good idea to treat the cells with either the plasmid or siRNA, and the drug at the same time?
Thanks in advance for your help!
This is impossible to answer without knowing the drug and method of transfection. It is potentially fine, but it's also possible that the drug could interact with your transfection reagents and inhibit the transfection. It's also possible that drug uptake could be higher in the presence of the transfection reagents.
If possible, I would suggest making stable knockdown cell lines using shRNA or CRISPRi, to avoid this issue completely.
Thank you Ciaran Daly
I'm using X-tremeGENE transfection reagent for plasmid, and DharmaFECT reagent for siRNA. The drug I'm using in cisplatin. Is there a database that shows potential interactions?
It was a very good point that you mentioned about higher cellular uptake of the drug. I should adjust the dose for sure.
- Badges
- Science method
- Similar topics
- Biological Science
- Ecology