I'm trying to think of a way to over-express RNA using a plasmid. Previously, we have clone a RNA of interest (e.g., lncRNA) into pcDNA3.1/Zeo (Thermo Fisher Scientific, #V860–20). Because there will be extra sequences in this plasmid vector (e.g., T7 promoter sequence), I'm looking for a way to have just the target RNA without any other sequence to be expressed.
Do you have a good idea?
The promoter sequence will not be a part of the RNA sequence. Design your construct such that the RNA has a G as +1 and the rest of the sequence will be your RNA. I have designed many RNAs that use a T7 promoter.... I don't have this issue of unwanted sequences.
This article may help your work
Article RNA Interference Using a Plasmid Construct Expressing Short-...
Dear Prof. Uchida,
Please, check L4440(Plasmid #1654) at Addgene. There, you can find other suitable options too.
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