Extended Data Fig. 9: PspCas13b lacks collateral activity in HEK 293T cells compared to RfxCas13d by fluorescence reporter assays.
Quantifications of (a) mcherry and (b) eGFP expression and (c) their representative fluorescence images in HEK 293T cells transfected with plasmids encoding mcherry, eGFP, PspCas13b and either non-targeting (NT) crRNA, wildtype (WT) PspCas13b crRNA12 or mutant (4–6 nt MSM) PspCas13b crRNA12; N = 3. Quantifications of (d) mcherry and (e) eGFP expression and (f) their representative fluorescence images in HEK 293 T cells transfected with plasmids encoding mcherry, eGFP, RfxCas13d and either non-targeting (NT) crRNA, wildtype (WT) RfxCas13d crRNA1 or mutant (19–21 nt MSM) RfxCas13d crRNA1; N = 3. (g) Quantifications of eGFP expression and its representative fluorescence images in HEK 293T cells transfected with plasmids encoding eGFP, RfxCas13d and either non-targeting (NT) crRNA or RfxCas13d crRNA1 (WT) targeting mcherry. 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). N is the number of independent biological replicates. Source data are provided as a Source data file.
