I transfected some cells with viral RNA with and without adding trypsin in the medium. After some days I detect more RNA in the wells were I added trypsin, so I am wondering if it is because the virus is replicating with the help of the trypsin or if it is just an artifact due to the inactivation of RNases by trypsin.
S-Protein and S-peptide, either separately or combined in the active enzyme RNase-S,
can be digested by trypsin, whereas the native enzyme, RNase-A, is not attacked.
Good question!
Do you have serum in your culture during transfection? The serum is abundant of anti-trypsins which inactivate trypsin immediately. Based on this, I think your observation is an artifact rather than an involvement of trypsin in protecting your RNA. May I know how many times you repeated your experiment? I suggest you to check with your cell number or confluency differences between the conditions, which can contribute to the differences in RNA production.
Please update me with more information so that I can understand better. Thanks
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