Extended Data Fig. 1: The silencing efficiency of various crRNAs is highly variable.
(a) Dose-response fitting of mCherry silencing data with non-targeting crRNA (NT) and 16 targeting crRNAs in Fig. 1d. (b) The relative IC50 values for 16 crRNAs targeting mCherry transcripts obtained from fitting in Fig. 1d. (c) PspCas13b expression in HEK 293 T cells expressing a targeting crRNA12 versus a non-targeting (NT) control crRNA. Data points in the graph are averages of normalized mean fluorescence from 4 representative fields of view imaged in N = 3. The data are represented in arbitrary units (A.U.). Errors are SEM and p-values of one-way Anova test are indicated (95% confidence interval). (d) Representative fluorescence microscopy images show the expression of PspCas13b in HEK 293 T cells expressing a targeting crRNA12 versus a non-targeting (NT) control crRNA; N = 3. N is the number of independent biological experiments. Source data are provided as a Source data file. Note: Calculating IC50 values for plasmids is not a common practice, given the challenges in quantifying the number of crRNA molecules being produced from a single plasmid in the cell. Thus, we call this value that estimates crRNA potency "IC50" a "relative IC50". The purpose of this ‘relative IC50’ is to compare the silencing activity of each crRNA in a quantitative manner.
