Normalization of miRNA activity profiles
This protocol is extracted from research article:
Numerical operations in living cells by programmable RNA devices
Sci Adv, Aug 21, 2019; DOI: 10.1126/sciadv.aax0835

Reporter expressions (eq. S2 in Supplementary Text) of 270 single-slot mRNAs in each cell type were determined from the compensated datasets of the flow cytometry. Notably, only one-third of miRNA species expressed in a cell repress the expression of their corresponding reporter mRNAs (24). This observation suggests that two-thirds of single-slot mRNAs used in this study were not affected by their corresponding miRNAs. Using reporter expressions in HeLa cells as a standard, the expressions of hmAG1, tagBFP, and hdKRed in each cell type were independently corrected according to simple linear regression without the intercept term to obtain bias-corrected expressions. Then, the mean and SD of the bias caused by differences in the reporter fluorescent proteins was heuristically set to 0.5 and 0.15, respectively.

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