The fold increase in activation of the UbiC promoter by dCas9-TA is calculated as the ratio between the normalized GFP intensity of CaT-A cells and CaT-B cells.
Below is an example calculation based on positive control MDA-MB-231-dCas9-VPH cells (see Additional Required Materials to obtain this cell line):
|Mean_GFP||Mean_RFP||Normalized GFP (GFP/RFP)|
|CaT-A Cells||266||126||266 / 126 = 2.111|
|CaT-B Cells||11.7||128||11.7 / 128 = 0.091|
dCas9-TA induced fold induction = 2.111 / 0.091 = 23.1 fold
As an additional negative control (optional), the parental cell line of the dCas9-TA cells can be transduced with CaT-A and CaT-B viruses. For cells without dCas9-TA, the fold activation should be close to 1.
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