Figure 5. Effect of targeted integration on homogeneous expression of GOI.

Targeted and random integrants isolated from the stable cell pool transfected with Cas9, sgRNA, and donor plasmid targeting COSMC locus were cultivated to assess (a) RFU and (b) relative specific growth rate (µ). Average mean intensity of mCherry fluorescence measured by imaging cell cytometer was used for RFU. The specific growth rate was calculated from a plot of viable cell concentration against culture time from day 0 to day 3. Each measured value was normalized by that of the clone number 1. Center lines show the medians; box limits indicate the 25th and 75th percentiles as determined by R software; whiskers extend 1.5 times the interquartile range from the 25th and 75th percentiles, outliers are represented by dots; data points are plotted as open circles. n = 41, 40 sample points. Each sample point represents average value of measurements at 3–5 different passages during cultivation for a month. (CV) coefficient of variation.