I am looking for a method to totally suppress RNA transcriptase in cell suspensions and see if any new strands of RNA are still elaborated as measured by uptake of tritiated nucleotides.
I think you can use siRNA to knock down key protein to regulate this process. like rna polymerase. I just guess.
there are certainly various reagents that block transcription by some RNA polymerases. Which organism or cells are you looking at?
Thank you for the helpful thread Joydeep. Michael, I want to totally inhibit transcription in macrophages collected by peritoneal lavage after injection of mineral oil. Bingxian, siRNA is not an option for me.
α-amanitin is an inhibitor of both RNA polymerases II and III in eukaryotic cells. However I don't think it inhibits RPO-I so I'm not sure if that works for your experiment or not.
Michael, any problem in using amanitin in combination with actinomicin D?
They work by different mechanisms so I imagine there is not a problem using both. But not sure if the amanit adds anything to act D or not. I imagine there are many pubs in the literature that address this.
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