In other words, how do you design a self-amplifying (replicon) RNA aiming to achieve the preferential expression of a particular mRNA? What other mRNAs need to be included in the replicon? Thanks.
Are you wanting to have a DNA plasmid that expresses the RNA preferentially? Or are you asking for a self-replicating RNA molecule? The first is easy, the second is much more difficult.
It will be the second case. The bacterial species to consider need to be enterobacteria. I understand RNA bacteriophages might be the right tools. I wonder if you would have any particular suggestions in this regard. Thank you so much for your engagement.
It has been done in E. coli at least, I'm not sure about other bacteria. You can make a cDNA copy of the phage genome (and introduce any modifications you might wish) and then express from a regulated promoter. Infectious phages are then replicated and produced.
Olsthoorn RC, Licis N, van Duin J. Leeway and constraints in the forced evolution of a regulatory RNA helix. The EMBO Journal. 1994;13(11):2660-2668.
I'm sure there are similar references for other phages but I don't know them off the top of my head.
Thanks so much Michael, I'll see if anyone else suggests a well characterized RNA coliphage anyway. Kind regards.
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