Targeting transcriptionally silent gene

[Pratiksha]

We have tried to target a transcriptionally silent gene with a reporter and antibiotic-resistant gene as a selection marker. We see confirmed integration of the cassette but no expression of the reporter upon differentiation. Is there a possibility that the antibiotic resistance gene is affecting the expression of the reporter?

[Janani]

Hi Pratiksha,

Thank you. We will need some more information about this experiment. Was selection performed? Could you maybe please share a map of the construct?

Thanks,
Janani

[Pratiksha]

Thank you for the reply. Our selection marker is puromycin resistant gene. The selection was performed for 10 days. Please find the attached vector map. This segment was cloned into a pBSK vector backbone.