I want express two mir-30 based shRNAs from a single transcript. I plan to modify the TRIPZ inducible system as following:
PolII inducible promoter - RFP - shRNA1 - shRNA2 (Clone in second shRNA).
Do I need a specific distance or sequence between shRNA 1 and 2? I assume RFP-shRNA1-shRNA2 will expressed as a single transcript. RFP will then be translated and shRNA1 and 2 will be processed from the transcript. Correct?
I cannot find any literature about this so please let me know if there is a good paper or if you have suggestions for the cloning.