Hi everyone,
I made plasmid construct, pSpecificPromoter-DIO-GeneA-P2A-mtdTomato. I transfected the plasmid into the neuronal cell line (lipofectamine2000, incubation time 48 hour). I found more than 10% of cells in transfected cells (approximately 2% of total cells) expressed tdTomato without Cre recombinase. DNA sequence of the construct is correct, tdTomato sequence was inverted.
Does anyone have this kind of experience?
Thank you.
Regards,
Yasuyuki
Yasuyuki Osanai
Hi everyone,
We found the reason. We found that GeneA posses promoter activity. When we changed the order of genes (tdTomato-P2A-GeneA) the leak expression was disappeared. I hope this experience help someones experiment.
Regards,
Yasu
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