I'm a student and I have to produce a cell line with a knockin for the NRF2 gene with GFP. I have to put a promotor in front of the GFP because the gene will be too far away from the promotor of NRF2. I want to add the CMV promotor to the GFP. To shorten my insert I want to remove the CMV enhancer. Will the promotor still work?
The CMV promoter does not require the enhancer region, but the enhancer is frequently included in many plasmids.
As an alternative, you might consider expressing NRF2 and GFP as a fusion protein, circumventing the need for anothe promoter. See:
Article A systematic analysis of Nrf2 pathway activation dynamics du...
Everything you can add to a plasmid, the better. Enhancers do increase the efficency. The CMV promotor, may still do, an enhancer increases the effectiveness of expression. The replication system clungs to the set of genes. Make sure there is some space between them (NRF2 and GFP). It is normally recommended some space.
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