Correlation of mRNA delivery timing and protein expression in lipid-based transfection

A. Reiser; D. Woschée; N. Mehrotra; R. Krzyszton; H. H. Strey; J. O. Rädler

The copyright holder for this preprint (which was not peer-reviewed) is the . https://doi.org/10.1101/607986 doi: bioRxiv preprint in Figure2B) are reliably identified by the model. In the supplement we provide further details about the accuracy of the parameter estimation. In particular, we compare our model to alternative approaches that were explored but that we found less suitable (see Supplementary Figure 5 ). We analyzed the data by an automated routine, which fitted each time course by the translation-maturation model. Since maturation rate and protein degradation were fixed, the routine yields the expression onset t0, the expression rate m0kTL, and the mRNA degradation rate δ. The distribution of single-cell parameters of onset times and expression rates are shown in Figure 2C . The onset time show a distribution that is slightly skewed and roughly described by a gamma function (see Supplementary Figure 3A ). Interestingly, the earliest observed onsets are as early as 10 min after incubation, while the later onset times extend too many hours. The average width of the onset time distribution measured as full width half maximum (FWHM) of the estimated distribution is about 2-3 hours. The expression rate is described by a log-normal distribution. The mRNA degradation rates δ also exhibit a distribution, an observation that will not be further discussed since it is outside the scope of this report (see Supplementary Figure 2 ).