Has anyone experienced loss of expression by plasmids after floxing their ORFs? It only happened with some promoters types. Secondary structures?

07 July 2016 0 4K Report

I have floxed an ORF+polyA directly under a neurons specific promoter in this configuration: Neu_promoter-loxP-ORF-polyA-loxP. This seems to impair the transcription since the gen (a reporter) is not longer expressed. This same configuration works perfectly under a regular CAG promoter. I wonder whether the distance between loxp sites allows the formation of secondary structures that may impede the function of the promotor.

I would appreciate whether anyone can share tips in this regards.

Thx!

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