I am trying to express m6A protein and a fluorescence one RFP in the piggyback vector.
I was thinking first of using the P2A to separate between the m6A and the RFP protein but then I was afraid of the residual P2A in the m6A if it would affect the localisation or the interaction at the C terminus, then the second choice would be IRIS, although the expression casste in the Piggybac itself has another IRIS followed by the selection marker
so the cassette would be as follows
m6A IRIS RFP IRIS puromycin
Is that ok or I would have a problem with the expression of the proteins
If there is please can you suggest other thoughts?